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Full text of "The human brain : histological and coarse methods of research : a manual for students and asylum medical officers"


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By W. BEYAN lewis, L.R.C.P. (Lond.) 





In these days of a voluminous scientific literatui'e the • 
appearance of every new manual demands some justifica- 
tion ; a genuine want can alone excuse additions to an 
already extensive series of handbooks of scientific methods 
of research. 

The onus 2J)-obandi of this want rests, of course, with the 
author,, and can in the present instance be readily met. 

Prompted by the editors of Brain, as well as by a 
personally-felt need for a concise summary of methods of 
examination of the nervous centres, the author contributed 
to the pages of that journal a series of articles on " Methods 
of Preparing, Demonstrating, and Examining Cerebral 
Structure in Health and Disease." 

An appeal thus made to what was regarded as a gene- 
rally prevailing want amongst asylum medical officers and 
students, was met with signs of cordial approval, and the 
frequently expressed wish for the re-appearance of these 
articles in a separate and convenient form. 

Such expressions of approval form the author's justi- 
fication for this Manual of Methods. 

West Riding Asylxtm, April, 1882. 



Coarse Examination of the Brain and its Membranes. 


The Duka Mater and Venous Sinuses. 


General Anatomical Features— General Appearance at the Vault — 
Examination of the larger Venous Sinuses — GenenU Appear- 
ance at the Base . . . . . . - . • • . • • . • 3-S 


The Aeachnoid and Pia Matek. 

General Anatomical Features — Coarse Examination of the Mem- 
branes.. .. .. .. .. .. .. .. ..iJ-H 


The Aetekial System of the Beain. 

Dissection — Arrangement of the Blood-supply in Circle of Wiliis— 
Superficial Examination of the Arterial Tunics — Capacity of 
the Arteries at the Base — Vessels of the Pia Mater — Nutrient 
Vessels at the Base . . . . . . . . . . . . 15-25 


Physical Properties of the Grey and "White Substance of 

the Brain. 


Conditions modifying Normal Consistence — Estimation of Textural 
Cohesion— Consistence of the Normal Brain — Reductions in 



Consistence: 1. Putrefactive Changes; 2. Indications of 
Softening; 3. White Softening of the Brain ; 4. Limited Foci 
of Softening; 5. YeUow Softening of the Brain; 6. Eed or 
Inflammatory Softening of the Brain ; 7. Simple OEdematous 
Conditions of the Brain— Augmented Consistence: 1. Super- 
ficial Scleroses; 2. Hypertrophy of Neuroglia; 3. Cicatricial 
Formations; 4. Disseminated Sclerosis; 5. Descending 
Sclerosis . . . . , . . . . . . . . . . . 45 

§ COLOUR 45 

Pallor: 1. Partial Mottling and Laminated Pallor; 2. General 
Diffused Pallor— Kedness : 1. Congested Zones ; 2. Patchy 
Kedness of Cortex ; 3. Puncta Vasculosa — Kesultsof Increased 
Intra-Cranial Pressure — Results of Increased Intra-Thoracic 
Pressure 55 

§ VOLUME . . . . 55 

Volumetric Methods for the Brain : 1. Dr. Hack Tuke's; 2. Other 
Methods ; 3. By Stevenson's Apparatus— Estimation of 
Cranial Capacity : 1. Dr. Hack Tuke's ; 2. Professor Flowers' ; 
3. Paraffin Method ; 4. Broca's Eesiilts .. ,. ..60 

§ WEIGHT 60 

Gravimetric Methods— Absolute Weight of Brain — Specific 
Gravity by : 1. Bucknill's Method ; 2. Sankey's Method ; 
3. Stevenson's Apparatus — Proportion of White and Grey 
Substance (Danilewsky) .. .. .. .. ..68 


Removal akd Dissection of the Beain. 









Minute Examination of the Brain. 


The Methods of Reseakch Conteasted 85 



Microtomes for Imbedding : 1. Stirling's ; 2. Ranvier's ; 3. Ruther- 
ford's; 4. Roy's; 5. Schiefferdecker's ; 6. For Large Sections 
— Microtomes for Freezing: 1. The Ether Freezing Micro- 
tome (Bevan Lewis) ; 2. The Ice and Salt Freezing Microtome 
(Rutherford) ; 3. The Quekett Club Freezing Microtome 
("Williams) 89-99 


Peepaeation of Haedened Beaijt. 

1. MiiUer's Fluid and Potassiimi Bichromate ; 2. Potassium 
Bichromate and Chromic Acid (Rutherford) ; 3. Iodized 
Spirit and Potassium Bichromate (Betz) ; 4. MiiUer's Fluid 
and Ammonium Bichromate (Hamilton) ; 5. Chromic Acid 
(Lockhart Clarke) .. .. ..105 




The Imbedding Mass— Section- Cutting — The Section-Blade— Sub- 
sequent Treatment of Sections — Sections through a Hemi- 
sphere — List of Apparatus Required ., .. .. ..114 


Hsematoxylin Staining: 1. The Dye; 2. The Staining Process; 
3. Notes on the Reaction of the Dye — Cai-mine Staining : 

1. The Dye; 2. The Staining Process; 3. Notes on the 
Reaction of the Dye — Picro-cannine Staining : 1 . The Dye ; 

2. The Staining Process ; 3. Notes on the Reaction of the Dye — 
Aniline Blue-black Staining : 1. The Dye ; 2. The Staining 
Process ; 3. Notes on the Reaction of the Dye — Osiuic Acid 
Staining (Exner' s Process) — On the Use of Compound Pigments : 
1. Ajiiline Picro-carminate ; 2. Picro-aniline Staining ; 

3. Osmium and Picro-carmine ; 4. Hsematoxylin with Aniline ; 

5. Eosin and Logwood ; 6. Picro-carmine and Iodine Green .. 131 





Staining the Cortex Cerebri — Staining the Cortex Cerebelli 
— Staining the Central Medulla and Ganglia— Staining the 
Pons, Medulla Oblongata, and Cord — List of Apparatus 
Required ... . . . . . . . • • • • • . . 1 3o 

Pkepaeation of Nekvous Tissue in the Fuesh State. 


The Freezing Method— Staining Fresh Sections of Cortex — 
Mounting for Permanent Preservation — Staining Fresh Sec- 
tions of Central Medulla 143 


Gerlach's Method for the Spinal Cord — Rapid Method for the 
Multipolar Cells of Cord — Rapid Method for the Ganglion Cells 
of the Cortex Cerebri : 1. First Stage, The Film; 2. Second 
Stage, Staining the FUm ; 3. Third Stage, Permanent Mount- 
ing — Examination of the Neuroglia: 1. Boll's Method; 
2. Ranvier's Method; 3. Mierzejowski's Method — Dissection of 
MeduUated Strands in the Fresh Brain — Remarks ni^on 
Dissociation of Nerve-CeUs .. .. .. .. ..1-49 


List of Keagents and Mounting Media. 

List of Hardening Reagents — Stock Solutions and Special Solu- 
tions .. .. .. .. .. .. .. .. ..153 

List of Staining Reagents — Stock Reagents and Special Solutions 155 
List of Macerating and Dissociating Reagents ,, ., ..158 

List of Media for the Mounting of Preparations . . . . . . 159 

List of Cements . . . . . . . . . . . . . . . . 159 



1. Diagram Illustrative of the Effects of Embolic Plugging (after 

Rindfleisch) . . . . . . . . . . . . . . . . 36 

2. Specific Gravity Test Apparatus . . . . . . . . . . 63 

3. Stevenson's Displacement Apparatus . . . . . . . . 65 

4. Convolutions of the Cerebnim as seen at the Vertex (after 


5. Convolutions of the Cerebrum, as seen at the Median Aspect 

(after Bischoff) 

6. Convolutions of the Cerebrum, as seen at the Base (after Bischoff) 

7. Convolutions of the Cerebrum, Lateral Aspect (after Bischoff) . . 

8 The Avithor's " Ether Freezing Microtome " (Original Model) . . 93 

<J. The Author's " Ether Freezing Microtome " (New Foi-m) . . 94 

10. Rutherford's Freezing Microtome .. .. .. .. .. 96 

1 1 . Williams' Freezing Microtome , . . . . . . . . . 98 




Important and necessary as are the minute investigations 
made by microscopic agency into the normal structure of the 
brain or its pathological deviations, we must guard ourselves 
from the very serious error of considering this method as 
essential or exclusively necessary in these studies. This 
error is a common one, and is too apparent to need mucli 
comment upon my part ; for all who are engaged in the 
prosecution of cerebral histology must have recognized the 
prevailing tendency to disregard the naked-eye appear- 
ances of the brain, or to consider them as of very secondary 
import compared with a minute investigation aided by the 
complex armamentarium of the microscopist. If any of 
my readers are possessed of this notion I would ask them 
at the outset to rid themselves immediately of so fallacious 
an idea as one which, if fostered, must prove a serious 
obstacle to the acquirement of that intimate acquaintance 
with the true significance of the varied appearances presented 
by normal and diseased tissues. The skilled obstetrician 
recognizes as an invaluable acquirement that tactus eruditus 
which a constant and intelligent employment of a special 


sense can alone confer ; and no less should the histologist 
endeavour to obtain a special visual tact, a highly refined and 
educated visual power, which can alone enable him to recog- 
nize by the unaided eye appearances which pass wholly 
unnoticed by the casual observer. I cannot too strongly 
insist upon this point, for he who would successfully study 
the morbid anatomy of the brain must, as in the morbid 
anatomy of other tissues, begin seriously to educate the eye 
to the coarse appearances presented to the unaided vision. 
The employment of a hand-lens of 2 to 4 inches focal 
length will prove of service here, the naked-eye appearance 
being contrasted with the magnified field, and the eye 
thus educated up to recognizing characters which without 
the aid of the lens were previously indefinite or unrecog- 
nizable. Nothing beyond repeated and energetic efforts in 
this direction will enlarge the area of the visual field and 
present to the mind the manifold appearances which consti- 
tute an unbroken whole, and wliich are absolutely necessary 
to a refined interpretation of the picture presented to the 
mind's eye. Repeatedly have I had occasion to observe that 
the student, after a full curriculum of hospital training — 
a training which should pre-eminently involve the high 
culture of the sense of sight, hearing, and touch — fails wholly 
to appreciate the most obvious abnormalities of the cerebro- 
spinal tissues, and this because he has neglected to tutor the 
eye so far as to learn what to look for, and how to look for it. 
If you place before him the brain of a case of chronic mania, 
without coarse lesion, and note his observations upon the 
appearances of a section across the hemisphere, it will be 
found that beyond a statement that the grey matter is pale, 
anoemie, and wasted, that the white matter is altered in con- 
sistence, and presents numerous coarse vessels, his untutored 
eye teaches him no more ; and should he be examined even 
for the grounds upon which these statements are made, a too 
evident vagueness mil be apparent — his ideas on relative 
depth of cortex or anoemic states of the brain and consistence 
of its tissue are ^ugcd by no mental standard, and are 


remarkable only for their indefiniteness. The practised eye 
of the histologist, however, sees far more : the relative depth 
of cortex in various regions ; the relation borne by this depth 
to normal standards ; the varied depths of the several laminoe 
of the cortex, their distinctness of outline, general and local 
vascularity, as well as the deviations in hue dej^endent upon 
fatty or upon pigmentary changes. The oedematous, degene- 
rated aspect of the medullary tracts, the presence of minute 
sclerosed patches, and a host of other morbid appearances, 
present to his mind a picture which the unpractised eye 
wholly fails to appreciate. To those who are liable to err in 
the direction pointed out, I now address myself in the hope 
of rendering their studies of the naked-eye appearance of 
healthy and diseased brain more inviting and instructive, by 
means of a few simple directions as to what appearances are 
to be sought and how they are to be looked for. 

A few fiu-ther statements on the object and scope of these 
articles may not prove amiss. 

I address myself almost exclusively to the student who, up 
to the present time, has devoted but little of his attention to 
cerebral pathology or to the histology of the central nervous 
system ; and more especially to asylum medical officers, 
whose opportunities for research in this field are so numerous, 
yet so sadly neglected. I shall endeavour here to place at 
the disposal of the student the more important, essential, 
and trustworthy information in this department which I find 
scattered promiscuously throughout an extensive literature. 
If amongst these gleanings my remarks should appear to the 
advanced student burdened by too much detail, my excuse 
must be that the primary object is to overcome the diffi- 
culties presented by these studies to the novice, difficulties 
which every expert observer has at the beginning to 





General Anatomical Features, — It is necessary here that 
the student should recall to mind certain anatomical details 
affecting the relationships of this membranous investment of 
the brain which have an important pathological significance, 
and in the first place note that : — 

1. The dura mater is a fibro-sevous membrane ; the outer 
sm-f ace being fibrous and rough, the inner being smooth and 
polished by a layer of epithehal cells constituting a parietal 

2. Cut off a small portion and float it in water. The rough 
pilose outer surface due to the numerous fibrous connections 
and vessels which unite it to the inner table of the skull 
becomes hereby very apparent in contrast to the smooth inner 

3. Next note that the dura mater is composed of Uco didinct 
layers, which, by their divergence, occasion the formation of 
the different venous sinuses ; whilst the inner layer, by its 
duplications, forms the various intra-cranial membranous 
partitions, the falx cerebri and cerebelli, as well as the 

4. Lastly, he must observe the close anatomical relation- 
ships between the lateral sinuses and the mastoid cells, and 
again between the superior petrosal sinuses and the internal 
ear. Caries of the petrous portion of the temporal bone and 
of its mastoid cells is so frequent an affection, that the con- 
tiguity to them of tliese venous sinuses is most important as 

1! 2 


likely to occasion not only localized pachymeningitis or 
inflammation of the opposed dura mater, but inflammation 
and thrombosis of the venous siniises, leading probably to 
metastatic deposits in the lung. 

5, The position of the vense Graleni, which receive the 
blood returning from the choroid plexuses and open into the 
straight sinus, is such that tumours or abscesses of the mid- 
lobe of the cerebellum would compress them. 

Upon removal of the skull-cap the student should proceed 
to investigate the condition of the dura mater after the follow- 
ing systematic manner : — 

1st. Note the general aspect of the membrane covering 
the hemispheres. 

2nd. Examine the several venous sinuses. 

3rd. Observe the condition of the dura mater in those 
regions at the base Avhere there exists a special 
proclivity to disease. 

General Appearance at the Vault. — A glance at the 
superficial aspect of the dura mater may reveal the presence of 
inflammatory products which are frequent here. If present, 
note their character, whether they are simple inflammatory 
exudates, capable of undergoing organization, or whether 
they be the results of suppui'ative inflammation. In the 
former case, observe how softened the membrane is, and how 
readily separable from the bone ; in the latter case the 
presence of pus will almost certainly lead to the detection of 
a carious state of the internal table and diploe of the skull, 
and a more or less sloughy aspect of the disintegrating mem- 
brane. The organizable • blastema maybe found in various 
stages of development, as loose areolar or thick tough fibrous 
tissue, or it may have formed bony plates which eventually 
unite with the cranial bones. Should dense fibrous bands be 
formed, adhesions betwixt the dura mater and bone occur, 
chiefly along the course of the sutures, or in localized islets 
over frontal or parietal bones, or so extensively that it is 


imj)Ossible to remove the skull-cap without at the same time 
including the dm-a mater, as their forcible separation would 
entail injury to the brain. 

If there be the above given indications on the sm'face of 
recent or of old inflammation, note the density, thickness, 
Sicollent appearcowe, and colour of the membrane at these parts. 
Recent inflammations involve much swelling, interstitial 
infiltration, softening and looseness of textiu'e, and a very 
red colour ; whilst the effects of an old inflammation are seen 
in a callous fibroid induration and thickening, often with 
bony concretions formed in the interstices of its textm-e, 
together with the adhesion to the inner table of the skull 
just alluded to, - 

The student must be prepared to make allowances for the 
general increase in thickness and firmer adhesion of the dura 
mater, which pertains to advanced age independent of in- 
flammatory changes. Superficial inspection will also enable 
him to detect attenuation of this membrane, such as results 
from the pressure of a hypertrophied brain, from the effusion 
of hydrocephalus or morbid growths. Local thinness of this 
membrane also may be due to the pressure of pacchionian 
bodies which perforate the dm-a mater frequently, and 
imbed themselves in fossae on the internal surface of the 

Extravasation of blood, forcing the membrane apart from 
the bone, may be frequently found, and its origin should be 
carefully sought. 

Examination of the larger Venous Sinuses. — A fold of 
dura mater close to the longitudinal sinus at its exposed frontal 
extremity must now be pinched up by a forceps, and the 
scalpel passed through it and do^vn into the longitudinal 
fissure of the brain, so as to divide the union betwixt the 
falx cerebri and crista galli. Introduce a curved bistoury 
into the anterior end of the longitudinal sinus, and carry the 
blade backwards to its occipital extremity so as to expose the 
sinus throughout its greater extent. The oblique orifices of 


the veins of the vertex which empty themselves into this 
sinus mil now be exposed. 

Note, 1st, The capacity of the sinus throughout. 

Note, 2n(i, The nature of its contents. 

Note, 3rd, The condition of its lining membrane. 

The incision through the frontal end of the falx cerebri 
should now be extended on either side through the dm\a 
mater backwards, upon a level with the sawn edge of the 
cranial bones. The anterior extremity of the falx being then 
seized with a forceps, should be drawn forcibly backwards 
from between the hemispheres, carefully dividing the great 
superficial veins where they open into the sinus, when the 
convolutionary aspect of the brain, covered by its soft mem- 
branes, will be exposed to view. At this stage of dissection 
the pia mater, arachnoid, and superficial aspect of the brain 
will require attention, and the method to be adopted will be 
detailed in the following chapter, to which the student must 
be referred. Continuing, however, our examination of the 
sinuses of the dura mater, it must be presumed that the brain 
has been removed, and the following procedure will then be 
necessary : — 

Lay open the larger venous sinuses at the base, commenc- 
ing at the torcular herophili behind, and carrying the blade 
down eacli lateral sinus to the foramen jugulare. Open up 
the straiglit sinus running into the torcular herophili in 
the base of the falx cerebri. Deal in like manner with the 
petrosal and cavernous sinuses. 

A close inspection of these sinuses should be instituted 
with the object of learning the natiu'e of their contents, and 
the morbid or healthy condition of the textm-es of these 
venous channels. The most frequent morbid conditions 
foimd here are those of phlebitis and thrombosis, the thi^ombus 
being usually the result of inflammation of the walls of the 

Thrombosis of the venous sinuses of the dura mater is so 
frequent and so important an affection, that the possibility of 


its occurrence should always suggest itself when examining 
this membrane. 

If a thrombus obstruct any one of these sinuses, note the 
more or less organized condition of the clot and its adhesion 
to the lining membrane of the sinus. 

Examine closely its apparent site of origin and extensions, 
the latter often reaching into the jugular veins. 

Next take into consideration the constitution of the clot, 
whether recent, organized and firm, or breaking up and pre- 
senting the appearance of a crumbling mass, which, dming 
life, would give rise to metastatic deposits in distant organs. 

The condition of the lining membrane and tissues of the 
sinus should next be noted. If inflamed, the inflammatory 
action should be traced to its origin, usually in a carious 
state of the cranial bones ; or by extension of simple inflam- 
mation of the dura mater in the neighbourhood of the sinus ; 
or less frequently, induced as a secondary resvdt of thrombosis. 
The student should carefidly study the effect of plugging of 
the different sinuses, and their proclivity to disease as a result 
of their varied anatomical relationships. 

1. Inflammation of the longitudinal sinus will be accom- 
panied by occlusion of the venous trunks from the convexity 
of the brain where they open into the sinus. 

2. If the straight sinus be occluded as the result of adhesive 
phlebitis, the student will remark that its effects in obstruct- 
ing the return of blood from the choroid plexus will be 
similar to what occiu-s when the venae Graleni are compressed 
by a tumour of the mid-lobe of the cerebellum. Effusion 
into the ventricles might be expected in these cases, and the 
enlargement of the head from this cause has been recognized 

3. If the lateral sinus be inflamed, we miglit expect, as 
a result, extension of phlebitis into the jugular veins, and 
plugging of the smaller sinuses or veins opening into it. In 
connection with this — note, that the small " emissary veins " 
(Santorini), which perforate the bone at the anterior extremity 
of the lateral sinus, and Avhich form a communication between 


tlie sinus and tlie veins on the outside of the skull at the back 
of the head, may become plugged by a thrombus producing 
a painful a3dema behind the ear, also recognized clinically by 

4. The position of the petrosal and lateral sinuses naturally 
exposes them to phlebitis and thrombosis. Lying on the 
petrous portion of the temporal bone, they are always liable 
to be implicated by extension of inflammation from the 
internal ear and caries of the temporal bone. 

5. Thrombosis of the cavernous sinus has been frequently 
noted. In connection with this condition the student must 
be prepared to note the pressure which will probably result 
to the carotid artery, which runs through this sinus invested 
by its lining membrane." He may also look for extension 
of inflammation of the coats of this sinus to the facial veins 
with which the ophthalmic vein communicates, noting the 
obstruction to the return of blood from the cavity of the 

6. Amongst other results of thi'ombosis of the cerebral 
veins and sinuses, will often be found punctiform hasmor- 
rhages into the substance of the brain. 

General Appearance at the Base. — Observe the con- 
dition of the dura mater at the base, where there exists a 
special proclivity to disease. The sites alluded to are : — 

1. Petrous portion of temporal bone. 

2. Ethmoidal plate. 

3. Parts adjoining superior cervical vertebrae. 

1 Quoted by Niemeyer in " Text-Book of Practical Medicine," vol. ii. 

2 This condition has been recognized clinically by a loud bruit heard on 
auscultation of the skull. Dowse, etc. Lancet. 



General Anatomical Features. — In proceeding to in- 
vestigate the healthy and morbid appearances of the arach- 
noid, the follomng anatomical facts will prove serviceable to 
the student. 

1. The arachnoid is usually regarded as consisting of a 
parietal and visceral layer like ordinary serous sacs, the 
former, however, being merely a layer of nucleated poly- 
gonal epithelium lining the inner surface of the dura 
mater ; the latter forming the far more substantial invest- 
ment of the convolutions of the brain separated from them 
by the pia mater. This closed sac is lubricated by a 
portion of the general cerebro-spinal fluid. 

2. The visceral layer of arachnoid does not dip into the 
sulci with the pia mater, but bridges them across, whilst in 
other regions it is widely separated from the pia mater so as 
to enclose between them extensive spaces. Thus, at the base, 
a delicate veil of arachnoid stretches across the pons and 
interpeduncular space as far as the optic chiasma ; a similar 
film of arachnoid closes in the fouiih ventricle, stretching 
across from the medidla to the cerebellum ; the arachnoid also 
does not pass down to the bottom of the longitudinal fissure, 
but spans across immediately beneath the falx cerebri ; hence 
a space is left here between the two inner membranes of the 
brain immediately above the corpus callosum. 

3. The various spaces alluded to above all communicate 
freely with one another, and with an extensive space around 


the whole length of the spinal cord betwixt the pia mater and 
arachnoid sac. The whole system of spaces is termed the 
subarachnoid space, and is filled by the greater bulk of the 
cerebro-spinal fluid. 

4. A space intervenes between the parietal layer of the 
arachnoid and the dm-a mater. It has been fully investigated 
and described by Axel Key and Ketzius/ and is called by 
these anatomists the sub-dura-mater space (subdiu-alraum). 
The student must therefore regard the membranous invest- 
ments of the brain as consisting of a lirm and immovable 
fibrous outer membrane, the dm'a mater forming also the 
periosteal lining of the cranial cavity ; of an intermediate 
serous sac or arachnoid whose visceral layer moves freely with 
the movements of the hemisphere ; and lastly, of a vascular 
membrane supporting the blood-supply of the brain — tlie pia 
mater, which closely lines the cortex, dipping into the sulci, 
and being prolonged into the ventricles. 

Between the various investments are fom* great cavities : — 

1. The subdui'alraum betwixt diu-a mater and the parietal 

2. The arachnoid cavity formed by the arachnoid sac. 

'3. The sub-arachnoid cavity betwixt visceral arachnoid and 
pia mater. 

4. The epi-cerebral space betAvixt pia mater and cortex. 

It must also be borne in mind that compensatory adjust- 
ments for lessened or decreased intra-cranial pressure are 
obtained by means of the cerebro-spinal fluid ; increase in the 
pressure, as in the increased amount of blood, gro-sHh of 
tumours, etc., being allowed for by escape of this fluid into 
the spinal sub-arachnoid space. In relation to this important 
subject it has been shown by Axel Key and Eetzius, that the 
pressm-e of the cerebro-spinal fluid is always higher than that 
of the venous blood, and its specific gravity lower ; hence its 

' "Studieu in der ADatomie des Ncrvensystems." Axel Key und Gustav 
Retzius. Stockholm, 1875. 


free mingling with the blood-currents by osmosis is much 

Coarse Examination of the Membranes. — The healthy 
and morbid appearance of the arachnoid should be subjected 
to close and critical examination. 

Upon reflecting the dura mater the contents of the arach- 
noid sac will be revealed, and in relation to this point we 
must note : — 

1. Variations in the amount of cerebro-spinal fluid here. 

2. Modified appearance of this fluid from admixtm-e with 
morbid products. 

3. Note especially extravasations of blood, and the age of 
these haemorrhages as revealed by the condition of the clot. 
It may be purely fluid blood, coagula, or a delicate film of 
fibrinous nature ; or it may be in various stages of organiza- 
tion. The various stages of the arachnoid cyst in relation to 
pachymeningitis have all been most graphically described. 

4. Note the superficial aspect of the visceral and parietal 
arachnoid as regards — 

a. Absence of clear, moist, glistening surface. 

h. Presence of morbid deposits, such as films of lymph of 
varied appearance and stage of organization ; as a 
delicate, greyish, mucus-like exudation, or a mem- 
brane possessing a certain amount of consistence, or 
flakes which are yellow and pimform, and more 
rarely fibroid patches ; or, again, the development 
of bony plates in the parietal layer. 

c. Adhesions between layers of arachnoid sac, such as 
occm' naturally by the development of the pac- 
chionian bodies. 

5. Note the anomalies of texture due to interstitial change 
and to deposits occurring here. The effect of frequent hyper- 

' Op. fit. 


reiiiia is seen in tlie opalescent, creamy white, or perfectly 
opaque appearance presented, especially along tlie course of 
the blood-vessels, and always seen after middle life to a 
greater or less extent. Hypertrophy of the textures, giving 
the membrane a tough and thick character, will result from 
similar causes. The membranes may be, however, soft, 
tumid, and swollen from cedema of its texture ; and where it 
bridges the sulci a solid gelatiniform aspect is often given it 
by the subjacent fluid. 

6. The relative amount of cerebro-si:)inal fluid in the sub- 
arachnoid space should next be taken into account. It should 
be collected, together with that which escapes from the arach- 
noid sac and ventricles, and carefully measiu'ed. The appear- 
ance of the arachnoid, when buoyed up by any undue 
amount of this fluid, should be noted before removal of the 
brain from the skull. It will be found that in these cases the 
arachnoid is widely separated from the pia mater by the sub- 
jacent fluid. The limpid character of this fluid may be 
tested by slitting up the arachnoid over a sulcus. 

So far our remarks chiefly apply to the arachnoid, and we 
will now turn our attention to the pia mater. The proximity 
of these two membranes to one another of course predisposes 
to their common implication in any morbid process, yet the 
pia mater is not by any means unusually involved without 
extension to the brain-substance, and still more frequently 
without extension to the arachnoid. In healthy states these 
membranes are very thin and delicate, and removed from 
the brain-surface with difficulty. "We should, therefore, 
note their proximity to each other over the summits of the 
gyi'i, alterations in the thicliness of the pia mater and in its 
toughness, infiltratimis of its texture, and the ease or difficulty 
experienced in its removal from the cortex. 

Thus it may be found widely separated from the arach- 
noid, the latter being buoyed up by sub-arachnoid effusion. 
In these cases the appearance of the fluid should be noted, 
especially as regards the presence of inflammatory exudates, 
as flakes of lymph, etc. 


If thickened in textm-e the alteration of the pia mater 
should be traced to its origin, whether this be simple serous 
infiltration or oedema conjoined with inflammatory material. 
Tortuosity and varicosity of the vessels will also be further 
indications of repeated congestions of this membrane. Espe- 
cial caution must be observed against drawing hasty conclu- 
sions regarding congestion of the membranes from the mere 
fulness of the blood-vessels and hypostasis.^ 

The student will find ample opportunity for studying the 
appearances presented by oedema and thickening of the soft 
membranes of the brain in cases of senile atrophy, general 
paralysis, and alcoholism. 

The presence of inflammatory material modifying the 
thickness and toughness of the membranes should lead to a 
study of the nature of the exudate, whether it appears as a 
tough exudate of plastic lymph more or less yellowish in 
hue, or more serous, sero-purulent, opaque, and less organ- 
izable material. 

With the presence of these more or less plastic exudates 
note the general infiltration of the membranes around with 
greyish, opaque serum. Such changes afford excellent 
opportunities for the study of the results of inflammatory 
action in loose areolar tissue. In connection ^vdth inflamma- 
tion of the pia mater — a condition which will frequently 
engage the attention of the student — he should also follow up 
his investigation of the nature of the exudate by examining 
also : — 

a. The direction taken by the morbid product, noting 

that this occiu-s almost invariably along the vas- 

cidar tracts, the coiu'se of the vessels being marked 

out by opacities. 

h. The extension to neighboming structm-es (brain, 

arachnoid, dura mater, and skull). 
c. The limitation of inflammation to s)naU areas (as 
when originating secondarily from caries of the 

' On this point see Niemeyer. Art. " Hyi^ersemia of Brain and its Mem- 
branes." "Text Book of Medicine," yoI. ii. 


cranial bones) ; or to the coiivexifij of the hemi- 
spheres (chiei3y accompanied by plastic products) ; 
or, lastly, characterized by the base of the brain 
being originally and chiefly involved (usually 
accompanied by aplastic and tubercular pro- 

Note attentively the greater or less facility of stripping 
the pia mater from the cortex. The membrane may be 
oeclematous, thickened, gelatinous, and most readily removed ; 
or, on the other hand, it may cling with greater tenacity or 
adhere so firmly that portions of the cortex tear away with it, 
leaving an eroded, worm-eaten aspect of the surface of the 

Attention should be directed to the strength of these adhe- 
sions, their implication of the summits of the gyri only, their 
localization over the convolutionary surface of ihe brain, and 
the coarseness of the blood-vessels entering the cortex at 
the sites of adhesion. 

Whenever the student is engaged with a case of menin- 
gitis, the morbid topography must be carefully studied. If 
basic, the veil of arachnoid extending from the optic chiasm 
to the pons must be examined and removed, noting imjolica- 
tion of the numerous vascular and fibrous twigs extending 
betwixt it and the subjacent pia : follow these results of 
inflammatory action up along the vessels of the fissures of 
Sylvius and the anterior cerebrals in the longitudinal fissure. 
Examine the regions just named for tubercular granulations, 
paying especial attention to the smaller blood-vessels, which 
may have tubercular masses in their sheaths, occluding more 
or less the calibre of the vessel. Should granulations appear 
around the blood-vessels, let them be removed and floated in 
water for more careful observation. 



Dissection. — After carefully noting the external appear- 
ance of the membranes covering the hemispheres both at the 
vertex and the base, the condition of the arterial system 
should be inquired into. The brain being placed with its 
base uppermost, the fine expanse of arachnoid which bridges 
across the inter-peduncidar space is first removed, and the 
great vessels forming the circle of Willis will then be ex- 
posed to view. Now separate the temporo-sphenoidal lobe 
from the adjacent frontal and parietal lobes by di\'iding 
the bridge of arachnoid extending between them across the 
Sylvian fissure. 

On gently drawing back the temporo-sphenoidal lobe the 
Sylvian branch of the middle cerebral artery will be observed 
running deeply in the fissure towards the upper extremity of 
the latter, giving off small branches in this coiu-se. The 
radiating gyri of the island of Reil (central lobe) vnW in 
most cases be only partially exposed, and it will be necessary 
to separate the opercuhmi, or that portion of the third or 
inferior frontal convolution which laps over the anterior 
portion of this lobe. The five or seven gyri of the island 
will now be seen radiating outwards like a fan and supporting 
the various branches deri-\-ed from the middle cerebral in its 
sulci. Split up the arachnoid which bridges across the longi- 
tudinal fissure anteriorly from one frontal lobe to the other. 
We shall thus have exposed the various branches of the circle 
of Willis as far as they can be seen at the base. Note first 
the arrangement of the blood- ve!<!<eh. 


The Circle of Willis. — A first glance at the arrange- 
ment of the great arteries at the base of the brain entering 
into the formation of the circle of "Willis cannot fail to 
impress upon the student's mind this important fact : there 
are here two great arterial systems more or less distinct and inde- 
pendent. In front lies the carotid system, supplying by far 
the greater bulk of the brain ; behind lies the vertebral system, 
distributed to the posterior and inferior regions of the cere- 
brum — an area small in comparison to the former. These 
two great arterial systems are united by the two posterior 
comnumicating arteries which connect each carotid with its 
corresponding posterior cerebral artery. 

Examine the posterior communicating arteries and note 
the remarkable smallness of their calibre, a fact which suffices 
to ensure us that the circulation in the carotid and vertebral 
systems is in the main distinct; whilst the arrangement 
allows of compensatory enlargement and a free communica- 
tion betwixt both systems where emboli, thrombi, or diseased 
textures obstruct the circulation of a main branch. 

Next observe the junction between the two anterior cere- 
brals deep in the longitudinal fissure by means of the cross 
branch, the anterior communicating artery. Raise the branch 
on the forceps and note its short length, small calibre, and 
right-angled direction, facts which teach us that, although in 
case of obstruction on the carotid side of the anterior cerebral, 
this branch, by dilating, may afford a satisfactory re-establish- 
ment of circulation over the area to which it is distributed, 
yet for the greater part the circulation betwixt the two anterior 
cerebrals is distinct. 

A little further consideration will teach us that the circu- 
lation of each hemisphere is by the above mechanism rendered 
wholly distinct and independent ; that the circulation of the 
mid-cerebral region of one side is wholly separated from that 
of the opposite hemisphere ; whilst the anterior cerebral areas 
are more closely associated through the medium of an anterior 
communicating branch. 

Lastly, through the medium of tlie posterior communi- 


eating artery (especially when we recall to mind the frequent 
enlargement of this vessel met with) there will be a far more 
ready communication betwixt the carotid and vertebral cir- 
culation than there can be betwixt the vascular apparatus of 
both hemispheres. We should pay especial attention, there- 
fore, to the following observations : — 

a. The almost comj)lete independence of hemispheric 

h. The very complete independence established betwixt 

the circulation of one middle cerebral and that of 

the other. 

c. The interdependence established betwixt the two 

anterior cerebral streams through the medium of 
an enlarged communicating branch. 

d. The possible admixture of the carotid and vertebral 

circulation of the same side through the medium 
of the posterior communicating — a condition very 
frequently established. 

The arrangement of the internal carotid ere it reaches the 
circle of Willis is one of interest and significance. Within 
its bony canal the tortuous sigmoid com'se taken by it is 
undoubtedly one means whereby the brain is protected from 
the results of the cardiac pulsations. The student will recall 
a similar tortuous course taken by the vertebral artery ere 
it enters the cranium. It has long been noticed that the 
middle cerebral is more readily blocked by emboli than the 
other branches, and that the area of its distribution shows 
especial proclivity to haemorrhage. Anatomical reasons, and 
the fact that this vessel lies " more directly in the way of 
strain fi'om the heart explains its frequent plugging and 
ruptui-e from disease " (Hughlings Jackson ^). Thus Prevost 
and Cotard found that tobacco seeds injected into a dog's 
carotid most often lodged in the middle cerebral. Again, the 
fact that the left carotid arises directly from the summit of 
the aortic arch, whilst the right, arising from the innominate, 

> Lancet, September 4, 1875. 




is inclined at an angle to the aortic current, is sufficient to 
explain the greater immunity from embolism experienced by 
the right carotid in its distribution to the brain. The right 
vertebral arises from the horizontal part of the subclavian, 
and is therefore also less subject to embolism than the left 
vertebral, which arises from the simimit of the ascending 
portion of the left subclavian. 

The Arterial Tunics. — Note the following points : — 

1. Colour and opacity. 

2. Relative toughness of coats. 

3. Tortuosity or kinks in the course of the vessel. 

4. Local bulgings or constrictions from diseases of texture. 

1. The arteries may be perfectly white in cases of slow 
and lingering death, and contain a decolourized clot extend- 
ing into its minute ramifications. On the other hand, they 
may be of a deep red hue from fluid blood, dark clots, and 
blood staining of the lining membrane. The vessels, again, 
may be thin and semi-transparent, or opaque from thicken- 
ing of the arterial tunics and morbid deposits. 

2. The toughness of the arterial tunics caji be tested later 
on after their removal. Their resistance to strain is an im- 
portant feature, and due attention should be paid to it by the 
student. The apparent thinness of a vessel should never 
be allowed to deceive him, as the thicker vessels are often 
most degenerated and least resistant to traction or expansion. 
Traction may be applied to the vessel held between two pairs 
of forceps, when an approximate idea of the breaking strain 
may be acquired, whilst the use of a conical gauge will 
enable him to note the various degrees of resistance these 
textures offer to expanding forces. 

3. Tortuosity and kinks in the smaller branches should be 
noted as suggestive of previous forcible distension from con- 
gestion. This is often apparent in the larger arteries ; thus, 
Quain has noticed frequent tortuosity of the internal carotid 
before it enters the carotid canal outside the skull in apoj^lec- 
tic subjects. 


4. Local biilgings may be clue to aneurismal dilatations, 
to the different forms of arteritis— atheroma, tubercle, 
syphilitic gummata, or to the impaction of a thi'ombus 
or embolus. The arteries at the base mil be found often 
exceedingly atheromatous, tortuous, knotty, and white ; the 
amount of atheromatous material occluding even the larger 
cerebral arteries, often converting the smaller branches into 
irregular knotty cords. The student should never infer 
from the aspect or feel of such a vessel that it is occluded, but 
he should always make a section across the mass of diseased 
tissue, when, if the vessel be permeable, the orifice is readily 

Note particularly the branches which are occluded. If 
the bulging be due to inflammatory swelling and exudation 
into the outer tunics of the artery, it will be observed that 
corresponding to the site of lesion the vessel is dilated — a 
condition due to paralysis of the muscular coat of the vessel, 
together with implication of the elastic outer timic. Make a 
section across the inflamed tissue, and observe how readily 
the elastic or outer coat may be stripped from the muscular 
by means of a forceps ; also how friable and easily separable 
are the muscular fibres of the media. The inner coat mil be 
probably deeply stained by htematin, or may be eroded and 
covered by an adherent clot immediately beneath the inflamed 
patch. When a clot appears to obstruct the calibre of a 
blood-vessel, the character of the clot should be taken into 
consideration, its fibrinous constitution, stage of organization, 
adhesion to vascular walls, its fomi, prolongations, and 
appearance of its section. The sheath and outer tunic of the 
arteries should be closely examined, especially in the neigh- 
bom^hood of the Sylvian fissure and island of Reil, for tuber- 
cular or syphilitic growths. Especial care must be taken to 
exactly note the arterial tunics involved, so as to discrimi- 
nate between ordinary endarteritis, residting in atheromatous 
degeneration, and the syphilitic node or gumma involving the 
outer coats, and the tubercular nodules involving the arterial 
sheaths, all of which lesions may give rise to blocking, con- 


traction of the cavity, and thrombosis. Such lesions should 
he examined microscopically. In all cases the above super- 
ficial examination should be supplemented by removal of 
short lengths of any diseased vessels, to be reserved for 
freezing and section-cutting, according to the methods 
detailed in the microscopic section. 

The vessels spreading over the island of Reil and up the 
Sylvian fissure should now be raised by passing a forceps 
beneath them, and with a clean sweep of the scalpel they 
should be all divided just where they turn over on to the 
superficial aspect of the brain ; this should be repeated on the 
opposite side. The two anterior cerebral arteries should now 
be divided at the genu of the corpus callosum, and dissected 
back to their origin from the carotids. The posterior cere- 
brals will be found running backM^ards round the crura 
cerebri under cover of a bridge of arachnoid. The posterior 
cerebrals, together with the three cerebellar branches, may 
be followed to a short distance and then divided. It will 
now be found that the circle of Willis is retained merely by 
the numerous minute nutritive branches, between the basilar 
artery and pons, and those from the anterior, middle, and 
posterior cerebrals, which pass through the anterior and pos- 
terior perforated sj)aces. Grently draw these vessels out by 
means of a forceps as far as possible, sever them close to the 
surface of the brain, and then float off the vessels of the base 
into a shallow dish or plate of water, arranging the branches 
in their relative positions. 

Capacity of the Arteries at the Base. — It may be 
found advisable in certain cases to test the relative capacity 
of the larger vessels at the base, and this may be obtained 
approximately by means of a small graduated conical gauge. 

In making these comparisons the student should bear in 
mind that the united areas of the branches equal very nearly 
the area of the trunk from which they originated, although 
their united diameters far exceed that of the latter. It has 
been shown by Paget's measurements that the equality 



between the area of the trunk and of its branches is not exactly 
maintained, the area sometimes increasing in the vessels of 
the upper extremities and head and neck, and diminishing 
in the lower extremities. Now, according to the mathe- 
matical law that the areas of circles are as the squares of 
their diameters, it will be necessary to contrast in our 
measurements the square of the diameter of the trunk with 
the sum of the squares of that of the branches arising from 
it. The student, therefore, should proceed in the following 
manner : — 

Let us suppose he wishes to estimate the relative capacity 
of the vessels at the base and theii' primary branches ; let 
him remove these vessels as already recommended, and float 
them out in a shallow vessel containing water. With a 
sharp pair of scissors cut across the vessel exactly at right 
angles to its direction, pass the graduated cone into it, and 
gently draw the vessel on until it is fully distended, but not 
stretched, by the gauge ; read off the diameter as shown on 
the gauge, and proceed in like manner mth the other vessels. 
Arrange the diameter and the squares in appropriate columns, 
opposite the names of the vessels, for future comparison. The 
following tables represent the average measurements of the 
various vessels at the base in forty-five cases of insanity I 
have investigated : — 


Vertebral Artery — Right 


Posterior Cerebral —Right 


Carotid — Right . . . 

„ Left .... 

Mid- Cerebral— Right . 

Left . . 

Anterior Cerebral — Right 





























Thus the sum of the square of the diameters of the 
anterior aud middle cerebrals of the right side was to that 
of the left side as 17-613 to 17-796. 

The square of the diameter of right and left anterior and 
middle cerebrals amounted in the aggregate to 35-409, both 
posterior cerebrals reaching only 13-764. 

Contrasting again the mid-cerebral supply of both sides 
with that of both anterior cerebrals, we find it represented 
by 20-414 against 14-995. 

Reference to the table of measurements will show that the 
posterior and anterior cerebral supply of both sides is almost 
exactly similar : thus, on the right side, we find 7-213 con- 
trasted with 7-55 ; and on the left side, 6-551 with 7-445. 

Another interesting feature in connection with these 
measurements is the great preponderance in areas of the two 
vertebrals over the basilar, which they form by their union. 
The two former have as the sum of the square of their 
diameters 22*38, the basilar only averaging 14-805 ; and in 
several cases I have found the basilar artery not more than 
one-half the area of the united vertebrals. 

It may assist the student to obtain a clearer idea of these 
areas if he represents them graphically as straight lines upon 
an enlarged scale. 

Vessels of the Pia Mater. — First note the general 
appearance and distribution of the vessels of the pia mater 
as they lie in situ. Observe the relative position taken by 
the veins and arteries, the former being large and sujierficial, 
the latter much smaller and concealed chiefly within the folds 
of pia mater, which dip into the sidci and are bridged across 
by arachnoid. The upper aspect being arachnoid, should, in 
health, present a smooth endothelial surface. Use a hand-lens 
in examining these vessels. The student should observe any 
fibrinous effusions, purulent exudates, or minute extravasa- 
tions along the course of the blood-vessels, the presence of 
atheroma, morbid growths — as gummata, tubercle, etc. If 
tuberculosis be suspected, examine the arteries deep in the 


sulci, and this with care, as tubercle along the course of the 
vessels is found with far greater difficulty here than at the 
base or up the Sylvian fissure. 

Next strip off a large portion of arachnoid and pia mater 
from the sm-face, and transfer it to a deep vessel containing 
water, the arachnoid siu"face being uppermost. Observe the 
velvety aspect of the lower surface, due to a fine pile of blood- 
vessels, and the very rich fringe of vessels along the inter- 
gyral folds, for the supply of the cortex deep within the sulci. 

Seize Avath a forceps one of these richly fringed folds and 
snip off a portion with a scissors, floating it on to a glass 
slide, with its vessels disentangled and spread out. Eemove 
also to another glass slide a portion of the membranes de- 
tached from the summit of a convolution, and study them by 
means of a hand-lens, both in reflected and transmitted light. 
Educate the eye in this manner to the recognition of the 
various healthy and morbid structures to be found here. 

For microscopic purposes, both these slides last mentioned 
should be reserved, whilst a small portion of brain covered 
by its membrane, and including two adjacent convolutions, 
should also be set aside. These will aiforcl subjects for a 
hrnVs-eye view of the surface under low powers of the micro- 
scope, such an examination being always insisted upon as 
especially valuable. The portion of brain must then be 
frozen and cut in such a direction that the sections so obtained 
represent both gyri with the intervening sulcus bridged over 
by arachnoid. We thus obtain very beautiful slides for fresh 
examination, which afford us the opportunity of minute 
examination of the cortex and the connections and relation- 
ships between it and its membranes. 

The large superficial veins at the vertex where they open 
into the longitudinal sinus, are occasionally found plugged 
by thrombi ; they shoidd therefore always be examined with 
the object of ascertaining the nature of their contents, espe- 
cially if there is evidence of inflammation of the dura mater 
or its sinuses, or of pm-ulent meningitis. 

The student should also take CA'ery opportunity of follow- 


ing up the arterial branches given off from the anterior, 
middle, and posterior cerebrals, in their course along the sulci, 
until he has familiarized himself with the exact supply of 
various regions and convolutions of the brain. It is only 
necessary to allude to the work of Duret and Charcot in this 
direction to indicate the importance of a minute knowledge 
of the arterial districts of the encephalon in order to fully 
appreciate numerous lesions met with in the brain. 

Nutrient Vessels at the Base. — We should devote 
attention to the numerous vascular tufts which arise from the 
large vessels at the base for the supply of the central ganglia 
and pons Varolii. Those which supply the basal ganglia 
are disposed into anterior and posterior groups — the former 
arising from the anterior cerebral, anterior communicating, 
and middle cerebral ; the latter taking origin from the bifur- 
cation of the basilar, often from an enlarged posterior com- 
municating, a further series arising from each posterior 
cerebral artery as it mnds around the crura cerebri. 
Roughly, it may be stated that the anterior group supplies 
the corpus striatum and anterior extremity of the thalamus 
opticus, whilst the posterior group is distributed chiefly to the 

Note, first, their remarkably fine calibre, considering their 
direct origin from main arterial trunks ; and secondly, observe 
that they are given off from these trunks at a right angle — a 
significant fact, as they can only be affected to a minimum 
degree by the propulsive wave of blood caused by each 
cardiac systole. They are filled, therefore, by a sustained 
lateral pressm*e. 

An examination of the nutrient supply to the pons will 
impress us with the follomng facts : — 

a. The nutrient twigs arising from the main artery are 
remarkably small, as in the other cases. 

h. They also arise at right angles from the basilar. 

c. The basilar is but slightly over half the capacity of 
both vertebrals, which supply it. 


The latter fact, which I have abeady demonstrated by 
measurement in our remarks upon arterial capacity, when 
taken in conj unction with the other two data, warrants us in 
assuming that these nutrient branches are kept constantly 
filled by high lateral pressure, and exhibit none of the 
phenomena of the pulse. I need not insist upon the beauty 
of this arrangement of means to end. 

In removing the vessels at the base, be careful not to tear 
these nutrient vessels short, but with gentle traction draw them 
out and cut them across with fine curved scissors ; float the 
circle of WilHs and its branches in a shallow dish of water, and 
examine the various nutrient groups. Snip off some of the 
anterior and posterior tufts, float them on to a slide, arrang- 
ing the branches, and examine by the naked eye as well as 
by microscopic aid. 

Wherever arterial degeneration is suspected, never miss the 
opportunity of thus closely inspecting the nutrient supply of 
the corpora striata, optic thalami, and pons Varolii. The 
naked eye will sufiice to reveal to us aneurismal dilatations 
along these branches, atheromatous degeneration, thrombi or 
embolic plugging, often explanatory of haemorrhages within 
the structure of the basal ganglia or pons. 



In the introductory chapter to his great work on Pathological 
Anatomy, Rokitansky has said : — " Just as there is a general 
and a special anatomy, physiology, pathology, so there must 
in like manner he a general and a special pathological 
anatomy. The former treats of anomalies of organization, 
the latter of the special anomalies of individual textures and 
organs.^ " It appears to me that in the examination of the 
diseased brain this natural classification must be kept in view. 
The student should be taught to appreciate the various general 
anomalies of organization as they present themselves in the 
brain ; and the methods to be adopted in estimating the 
degree, extent, and significance of such physical deviations. 
It is, of course, not our function to detail the various diseases 
of texture to which the brain is subject, but rather to place 
the student in possession of those indications of healthy and 
diseased tissues which are presented by alterations in the 
physical properties of grey and white matter. The elements 
of a physical diagnosis, so to speak, are placed at his disposal, 
and hints thrown out for his guidance, where it is thought 
a wrong construction might be attached to the physical 

It will be found convenient, however, to illustrate the more 
profound anomalies in consistence, colour, specific weight, and 
other physical qualities by reference to some of the i^pecial 
diseases of texture to which the nervous centres are liable, and 

' "Pathological Anatomy," Sydenham Soc. vol. i. 


which may serve as typical eases wherein the student may 
acquire a practical acquaintance mth the methods of pliysical 

The fundamental physical properties of the nervous tissues 
with which it is necessary we should acquaint ourselves are 
briefly as follows : — Consistence, colour, volume, and weight, 
both absolute and specific. Each in turn will now claim our 
close attention. 


The firmness, solidity, or consistence of a textm^e depends 
upon the cohesive force exerted by the individual elements 
of its mass, and to which is due, on the one hand, the varying 
degrees of toughness, hardness, and resistance, or, on the other 
hand, of softening and friability and liquefaction, to which 
they are liable. The consistence of a compound tissue so 
complex as the brain is necessarily subject to very great 
variations, since its individual constituents vary amongst 
themselves greatly in their physical properties. An organ 
composed of nerve-tubuli, nerve-cells, of a connective frame- 
work, and an elaborate system of arteries and veins, the whole 
pervaded by nutrient fluid, must be subjected to great modi- 
fications in consistence due to alterations in the relative pro- 
portions of one or other of the constituents. Hence it is 
that we get such variety in the structural firmness of brain- 
tissue, not only in different animals and in the brain of man, 
but in different regions of the same brain. It is this textiu^al 
difference betwixt grey and white matter which explains 
their varying degrees of consistence, the same explana- 
tion pertains to the firmness of the occipital as contrasted 
with the frontal lobe, and of the central grey ganglia as con- 
trasted ^\ith the grey envelope or cortex of the cerebrum. 
The grey cortex, consisting of a vast assemblage of nerve- 
cells and their protoplasmic extensions, imbedded in a most 
delicate web of connective tissue, with a complicated vascular 
apparatus, has a far lower degree of consistence than the 


white matter which owes its solidity to large meduUated 
nerve-fibres and an abundant and coarser connective matrix. 

A moment's consideration will suffice to indicate that the 
forces which in disease modify textural cohesion must come 
from without or from within the structural elements ; in other 
words, the tissue elements may be forced asunder by fluids 
pervading the texture and by the intrusion of adventitious 
bodies, or by nutritive changes in the elements themselves, 
whereby a quantitative or qualitative transformation may be 

Conditions modifying Normal Consistence. — 1. The 
forcible infiltration of nervous texture by serous fluid, by 
plastic exudates, or by blood, will in the first place tend to 
destroy all textural cohesion, and consequently produce reduc- 
tions in consistence ; whilst further changes in the effused 
blastemata due to organization will result in increased firm- 
ness or indiu-ation. 

2. Chemical changes in the constitution of the individual 
elements, according to their nature, may tend to produce 
increase or reduction of consistence, e.g., fatty, amyloid, and 
calcareous degenerations, the nutritive anomalies due to 
inflammation, decomposition of the stnictural elements due 
to putrefactive changes. 

3. Apart from the above qualitative anomalies of nutrition, 
we must not overlook the quantitative element which some- 
times exists alone or in combination with the former. 

Such are the forms of hypertrophy and atrophy due to an 
increase in size and number of constituent elements on the one 
side, and on the other, in the decrease in size, wasting, and 
diminution in the number of the tissue elements. 

The student must guard against the fallacy of regarding 
density and consistence of a tissue as in any way mutually 
convertible terms, for although in a large number of cases 
increased density may co-exist with increased consistence of 
the brain (sclerosis, hypertrophy, etc.), yet conditions occur 
where with increase of density there is really a diminution 


in consistence. Thus, in inflammatory conditions of the 
brain-tissue where plastic exudates have forcibly reduced the 
cohesion of the mass, the specific \iceiglit of the part is notably 
increased. Hence the specific gravity of cerebral tissue will 
afford us no exact and reliable guide as to degrees of con- 
sistence ; and, in fact, we have no more exact gauge of 
consistence of texture than the rough-and-ready methods 
afforded by the sense of sight and touch. 

Estimation of Textural Cohesion. — 1. By the eye we 
note deviations from the natural compactness of an organ, or 
the maintenance of its normal contour. Contrast the brain 
of a senile dement or of a general paralytic with the compact 
firm brain of epilepsy ; or the same organ in warm weather 
and in a state of incipient decomposition with the recent brain 
removed from the skull in cold frosty weather. 

2. By the sense of touch note the relative resistance to 
pressure and the compressible and lacerable character of the 
tissues. Especially note this with respect to the central com- 
missural tracts, basal ganglia, fornix, and septum lucidum. 

3. By its resistance to a graduated force, such as the rough 
gauge afforded by a stream of water falling from a variable 
height (applicable only to cases of much reduced consistence). 
Any case of white or inflammatory softening may thus be 

4. By its resistance to section (applicable to the slighter 
degrees of reduction and all degrees of increased consistence). 
Contrast the resistance to section presented by the medulla, 
pons, cerebellum, cerebral hemispheres, and ganglia at base. 

Observe that we gain by these means not only a satisfac- 
tory gauge of consistence, but that the methods vary in their 
relative value in different cases. We should take every 
opportimity of rendering oiu-selves perfectly familiar with 
the general consistence of healthy brain prior to post-moi-tem 
change. Let us remove a normal brain for careful study, 
and note the following points : — 


The Normal Brain. — Observe that upon removal from 
the cranial cavity it preserves its normal contour, although it 
has lost its natural support — the cranial bones. It is plump, 
rounded, and compact, giving a general impression of firmness 
and solidity, ere we gauge its consistence further by handling. 
The hemispheres are closely approximated by means of the 
strong commissural band — the corpus callosmn, showing no 
tendency to fall apart from rupture of the commissui-e, such 
as is so frequently seen in the degenerated and diseased brain. 
Upon separating the cerebral hemispheres with the hand the 
corpus callosum appears intact, and offers fair resistance to 
the blade when dividing the hemispheres along this course. 
The individual lobes maintain their characteristic forms, 
their salient margins, and relative positions ; each con- 
volution remains firm, plump, and in close contact with 
its neighboui% whilst the arachnoid investment gives a 
uniform, smooth, glossy aspect to the consolidated organ 

A section across the centrum ovale reveals a similar con- 
dition — observe the general solidity of the parts ; how per- 
fectly relative positions are maintained ; the absence of any 
gaping of the sulci ; observe also that the white substance 
cuts with a clean section, and shows no tendency to cling to 
the blade. Next open up the lateral ventricles, and observe 
the rounded firm aspect of the intraventricular portion of 
the corpus striatum and thalamus. Examine the fornix and 
septum lucidum — parts especially prone to softening in certain 
affections of the brain. Pay special attention to the relative 
consistence and size of the corpora quadi'igemina, the cerebral 
peduncles, the pons and medulla. Now we must be prepared 
to meet with all varieties of consistence betwixt the above- 
described normal firmness of health and extreme alterations 
produced as the result of disease. The consistence, as we 
shall illustrate further on, may be so far reduced that the 
cerebral substance may be perfectly diffluent, and may be 
poured away like thin cream ; or, on the other hand, it may 
possess the firmness, aspect, and character of the hard-boiled 


white of egg ; or even cause the knife to creak in cutting 
through patches of almost fibro-cartilaginous induration. 

Reductions in Consistence. — With the object of ren- 
dering ourselves familiar with reductions in consistence, 
let us consider briefly the appearance and significance of those 
abnormal conditions of cerebral tissue which lead to softening, 
dwelling only upon those which are most likely to present 
themselves frequently to om- examination. The conditions 
best illustrative of reduced consistence are the following : — 

Putrefactive changes. 

Softening as the result of disease. 

a. White softening. 

b. Yellow or gelatinous softening. 

c. Red or inflammatory softening. 
Simple oedematous conditions. 

1. Putrefactive Changes. — ^Now these changes in the 
brain occur sooner or later after death, according to the tem- 
peratm-e andhiunidityof the surrounding atmosphere, modified, 
of course, by the morbid conditions of its texture and con- 
tained fluid. It is on this account that the report of an 
examination of the brain should invariably be accompanied 
by a statement of the number of Jwurs offer death at which 
the examination was made, the average temperature of the 
post-mortem- room, and the condition of the atmosphere ns to 
humidity. Fremy, in an elaborate research in 1841 ("Ann. 
Chim," 2, 463), arrived at the conclusion that the ordinary 
form of softening of the brain was analogous in its production 
to the putrefactive process occurring post-mortem ; an opinion 
which has not been supported by later researches into the 
chemistry of the brain. We shoidd learn to discriminate 
between the softening due to this cause, and that due to 
morbid alteration of texture. We are guided here by observ- 
ing that the softening is general throughout the encephalon, 
and attended by much blood-staining from diapedesis, by the 
evolution of offensive gas, and the presence of the latter 


within the blood-vessels and beneath the membranes. We 
must also take into account the absence of any morbid con- 
dition tending to produce softening, and the conditions, 
atmospheric and otherwise, to which the brain has been exposed 
since death. It is essential, also, to keep in mind the fact 
that various diseases predispose to early putrefactive changes, 
whilst the mode of death may expedite or defer the same. 

To retard putrefactive changes when the brain cannot be 
immediately examined, it shoidd be placed in a cold room 
with a damp cloth thrown over the hemisphere to prevent 
desiccation of the outer cortical layers ; or better still, it may 
be enclosed within an ice safe. The student can readily 
extemporize for his own use such a safe, by placing the brain 
in a jar or box which fits into a larger box, leaving a space of 
an inch and a half around between the outer and inner box, 
into which ice is packed. The outer vessels must be thickly and 
completely enclosed in felt. Such a contrivance he will find 
of great service in the summer months, when it is difficult to 
keep anatomical subjects fresh for many hours together. 
Portions of brain reserved for microscopic examination after 
hardening should be transferred immediately to methylated 
spirits or Mliller's fluid, or better still to methylated 
spirits colom-ed of a dark sherry tint by tincture of iodine. 
For the fresh methods of examination, we must trust to the 
ice safe for the preservation of om- material, should any delay 
occur, preservative fluids being scarcely admissible. As 
regards the latter, however, the strong solution of acetate 
of ammonium recommended by Sankey ^ is perhaps the best 
which can be used (sp. gr. r040). 

2. Indications of Softening. — A general reduction in 
the consistence of the brain will usuoUy reveal itself at first 
sight by a mere superficial glance. The whole brain assmnes 
an unusually flattened squat appearance, and the hemispheres 
diverge. The student will find the former due to extension 

1 " New Process for Examining Brain Structure," West Riding Asylum 
Reports, vol. v. p. 192. 


of softeuiDg to the central portions of the hemispheres which 
support in the normal state the convolutionary sm*face folded 
over them, whilst he observes the commissural band connect- 
ing the hemispheres (corpus callosiun) tends to split asunder 
in a longitudinal direction — i.e., across the course of the great 
bulk of its softened fibres. Proceeding to handle the brain, 
he finds the convolutions flabby to the touch and presenting 
less resistance to pressm^e — the whole brain is less able to 
resist the ordinary force of gravity, and is therefore strongly 
contrasted with the plump, erect, and compact aspect of the 
healthy organ. Let the student examine the brain of an 
advanced general paralytic, and he will find this condition 
well represented. Cut across the hemisphere of such a brain 
so as to expose the centrum ovale, and it will be observed that 
the brain-tissue clings to the blade with unusual tenacity 
unless the latter be kept constantly wet by water or spirit. 
As a result, the cerebral tissues, both grey and white, are 
lacerated and tear away in slireds, leaving an unmistakable 
softened, rottenecl aspect of the surface of the section. This 
tearing away of brain-tissue is apt to give one a false idea of 
the couiparatire coarseness of texfure ; thus, in the brain of a 
general paralytic the adherent, thickened membrane and 
softened cortex cannot be cut without leaving a coarse 
irregular sm'face, unless the precaution be taken of using a 
very sharp blade, and keeping its siu'face constantly wet. 
Let us now examine more carefully the special form of soften- 
ing kno^vn as white softening. 

3. White Softening of the Brain. — AVe ma}^ meet 
with this condition as one generally diffused through a hemi- 
sphere or in patches limited to a convolution, the area of 
softening being often no larger than a pea, although more 
extensive tracts are usually involved. 

In the fu'st place, suppose we have before us a brain in 
which the greater part has thus sirffered. The affected 
hemisphere presents to the touch a soft, boggy feel, sometimes 
communicating an almost tremulous, fluctuating sensation to 



the fingers if the disintegration be extreme. The brain will 
be with difficulty removed, the disintegrated tissue tending 
to burst through the softened walls which confine it, and 
more especially is this the case in the neighbourhood of the 
anterior perforated space and commencement of the Syhian 
fissm-e. All unnecessary manipulation of such a brain should 
be avoided, as it is attended with danger to the textural 
continuity of internal parts, which should also be examined 
at as early a stage as possible. Observe the lateral divergence 
of the hemispheres and their flattened aspect above. By 
gently drawing the hemispheres apart and introducing a 
blade into the longitudinal fissiu-e, a section may be made 
outwards across the affected hemisphere, and thus removing 
the vault we find the centrum semi-ovale occupied by a mass 
of broken-do"v\Ti medulla — all normal cohesion being destroyed 
by the presence of a large quantity of interstitial serous 
effusion. The disintegrated substance will probably have 
the appearance of soft watery or creamy pap, readily raised 
upon the blade, or it may be absolutely diffluent, flowing 
away as soon as the resistance ofPered by the confining grey 
cortex is overcome. It may, however, retain a certain amount 
of cohesion, yet break up readily and wash away on placing 
it beneath a stream of water. The ganglia at the base may 
be implicated, and the septum pellucidum, grey commissure, 
fornix, and corpus callosum will generally be found softened 
or even broken down, the ventricle containing more or less 
serous fluid. Now such a brain as we have just been con- 
sidering will be met with in acute hydrocephalus, in tlii-om- 
bosis, or in embolic plugging of the main cerebral arteries. 
With such a case before him the student should pay attention 
to the following points : — 

a. Degree of softening — 

Does it resist a stream of water ? 

Does it render a stream of water milky or break 

down readily under it ? 
Is it from the first diffluent ? 

h. Extent of area involved. 


c. Q^dematous condition of the parts involved. 

d. Turbidity or clearness and colour of effused fluid in the 

In noting the degree of sofleniiuj learn to appreciate how 
very short a period is requisite, after arrest of blood-supply, 
for the production of extensive nutritive changes. 

With what object do w^e endeavour accurately to map out 
the extmt of the destroying lesion ? Oui' object is to detect 
that portion of the vascular apparatus involved if the soften- 
ing be due, as it most frequently is, to plugging of a large 
branch. This, the more frequent source of extensive white 
softening, requires, therefore, for its full comprehension, an 
acquaintance with the vascular areas of the hrain. 

On noting the appearance of the serous ep)ision in the 
ventricles, so often accompanying white softening, the student 
will be guided towards concluding as to the presence or not 
of inflammatory action in the meninges or in the substance of 
the brain itself. Thus, if the fluid be more or less turbid and 
discolom^ed (as tlie result of a plastic exudate and macerated 
cerebral substance), we may reasonably suspect an inflam- 
matory condition, and expect to find, upon microscopic 
examination, exudation corpuscles, compound cells of Gluge, 
and other products of inflammation. Such are the conditions 
found in acute hj'drocephalus, distinguishing it from a purely 
non-inflammatory form of hydrocephalus, which occurs where 
the fluid, though perhaps slightly tm'bicl from broken-down 
cerebral tissue and shreds from the macerated lining mem- 
brane of the ventricles, presents no inflammatory material 
amongst the debris. 

4. Limited Foci of Softening. — Acquaintance with 
the peculiar arrangement of the cerebral blood-vessels soon 
leads the student to infer that very minute tracts of softening- 
may result as the effect of thrombosis or embolism, the area 
involved being dependent upon the size of the clot and the 
site of its arrest or formation. It is necessary that the 
mechanism of white softening of cerebral tissue, as a result 

n 2 



of thrombosis or embolism, be thoroughly imderstood. The 
process may be best elucidated by the diagram below. 

a a 


Fig. 1. — Diagram illustrative of the Effects of Embolic Plugging 

(after Rindfleisch) . 
an. Portion deprived of its blood-supply Ijy the embolus. A, Artery. V. Vein filled 
with blood-clot. The airows indicate the collateral channels -which lead to a hyperfemic 
zone around the occluded vessels. 

Beyond the obstructed artery is the wedge-shaped area of 
its distribution, now anfemic and consequently deprived of 
its functional power. Below the embolus are seen swollen 
l)ranches, which tend to establish a collateral circulation. If 
this fails, we get as a result engorgement of the latter vessels, 
and a congestive vascular zone surroimding the wedge-shaped 
area. The tissue here becomes swollen and oedematous, and 
minute haemorrhages are apt to occur, whilst the whole 
central and peripheral texture becomes broken up by the 
effusion, and a true necrosis occurs of the tissue forming the 
area of distribution of the nutrient branch which has been 

All inflammatory foci have, as a result, the production of a 
similar congestion and aidema of the surrounding texture, 
accompanied by white softening ; and as new growths or 
tumovu-s are frequently the site of such inflammatory states, 
we may find them imbedded in a cavity containing broken- 
do\\Ti tissue. 

In addition, therefore, to the appearances which we have to 
look for in white softening of the brain, let us also note — 

Any obstruction to the vascular supply by thrombi, emboli. 


or adventitious products pressing upon tlie vessels from 

The establishment of a collateral circulation. 

The presence of inflammatory foci, to which the white 
softening is secondary. 

5. Yellow Softening. — In the course of pathological 
studies we frequently meet with appearances which, whilst 
significant of cerebral softening, differ much from that just 
described. We find frequently in the white substances of 
the hemispheres, less frequently in the cerebellum, and 
rarely on the convolutionary siu'face of the cerebrum, a focus 
of softened tissue, varying in size, but scarcely ever larger 
than a hen's egg, and characterized by its bright straw-yellow 
coloixr and soft gelatinous consistence. Pressure causes a 
yellowish fluid to exude, and when cut across it is found to 
be somewhat sharply defined from the pale swollen and 
ojdematous tissue around. We recognize in these softened 
areas the lesion termed " yellow softening of the brain," 

Fmiher examination reveals the fact that these spots may 
be primary, or, on the other hand, secondarily induced around 
adventitious products, such as tubercle, cancer, cysts, haemor- 
rhages, or around a patch of inflamed tissue. If secondary 
to adventitious growths, there is, as Rokitansky points out, 
an intermediate zone of red softening betwixt the growth and 
the yellow softened exterior. 

It is important that these patches should not mislead the 
student. He must from the outset regard them as identical 
in natm-e ^vith the first form of softening described, the 
difference in colour being due, according to some, to altered 
blood-pigment, but according to Rokitansky, to a peculiar 
chemico-pathological process. 

The following facts are to be noted : — 

a. The fluid is usually intensely acid. 

h. There is little or no vascularity smTOunding the patch. 

c. Every degree of tint may be foimd, from white 

' Plastic exudates, tubercle, .sypliilitic gummata. 


softening up to tlie bright yellow of typical yellow 

d. Colour differs from the rusty or ochre-yellow tint 

seen in old apoplectic cavities, etc. 
c. Microscopic examination fails to reveal inflammatory 


6. Red or Inflammatory Softening. — Profound altera- 
tions in consistence may thus attend inflammation of the 
nervous structures, and we find little difficulty in recognizing 
foci of inflammatory softening. "We occasionally meet with 
portions of acutely inflamed tissue in the medidlary strands 
of the cerebrum, varying in size from a hazel-niit to that of 
an orange, or still larger ; yet by far the more frequent site 
of such inflammatory foci will bo the grey cortex of the cere- 
brum and the basal ganglia. 

Examine a brain which presents a focus of inflammatory 
softening in the medulla of one of its hemispheres. Note the 
following facts : — The softenitig of texture is accompanied by 
modifications of colour and general moisture. Thus the increase 
in vascularity gives the affected part a streaky red aspect, 
often profusely besprinkled by puncta vasculosa ; the oedema- 
tous infiltration of texture is recognized, in its swollen and 
^noist aspect. Where the engorgement has reached a high 
degree, note the numerous minute extravasations of blood, the 
streaked or punctated capillary haemorrhages. 

Now all the above characters may be developed in the 
course of extreme congestion. What impresses upon the 
affected part the stamp of inflammatory action is the presence 
of exudates, which modify the above-described appearances 
in two directions : — 

a. The dark red becomes of a paler and more uniform 

hue, whilst exudations of lymph still further modify 
the aspect. 

b. Alterations in consistence occur; rapid softening or 

solutions of textural continuity varying with the 
plasticity of the exudates. 


Observe, therefore, that the essential features of red inflam- 
matory softening are profound alterations of consistence, 
associated with the pouring out of inflammatory exudations. 

In advanced stages of red inflammatory softening, the 
student will, therefore, find the tissues broken down into a red- 
dish pulp, or having a brownish or rusty aspect, washing away 
freely when held beneath a stream of water. He will also 
note the results of swelling from congestion and oedema, viz., 
the flattening of the convolutions of the hemispheres from 
pressm^e against the internal table of the skull, and a well- 
marked prominence of the surface of a section of the inflamed 
and oedematous textures. 

Examine a patch of inflammatory softening in the coi-tex, 
and note : — 

Inflammatory foci situated in the cortex will implicate by 
contiguity the superimjjosed membranes. Observe in these 
cases that the grey matter is more deeply coloured, and is 
still more swollen, moist, and softened than when the white 
matter is involved. The far greater vascularity and the 
naturally looser texture of the grey matter account for this 
difference. Next direct attention to the state of the mem- 
branes. The pia mater cannot be removed without peeling 
off with it a layer of the softened cortex, whilst the meshes of 
the membrane are found inflltrated with inflammatory pro- 
ducts, and are both thickened and oedematous. As regards 
these apparent adhesions of pia mater, observe that the brain 
substance may tear away with the removal of the pia mater 
from two causes : — 

1st. Mere unnatural softening and loss of textural cohesion 
of the grey matter, without any true adhesion to the mem- 
branes, as in early inflammatory and congestive stages, etc. 

2nd. Actual adhesion, inflammatory in origin, may be the 
cause of this tearing of the cortex. 

How are we to distinguish the laeerable cortex in the 
former case from the genuine inflammatory adhesions of the 
membranes ? Note, in the former case, the surface left by the 
in-egular sln-eds torn off is soft, pappy, and uniformly smooth 


or liomogeneous ; in the latter case, tlie surface is studded 
with numerous perforations — the apertures of perivascular 
channels from which the blood-vessels have been withdi'awn. 
These channels will be seen much dilated from the prior 
engorgement of the blood-vessels, and the surface generally 
has a peculiar worm-eaten appearance. 

FALLACY TO BE AVOIDED.— We imxst not regard 
all cases of encephalitis as necessarily attended by injection 
and discoloration of tissue. The larger number of cases, in 
fact, show little or no discoloration ; and even where softening 
has advanced to an extreme degree, the naked eye fails to 
appreciate in the uniformly dull white pulj^y material its 
inflammatory origin. 

What criterion have we here for determining the nature of 
the lesion ? Resource must be had to the microscojie and 
specific gravity bulbs. 

The niicroscope shows us nuclei, nucleoli, pigment, and 
compound granule-cells amongst broken-down cerebral tissue 
infiltrated with leucocytes and effused lymph along the 
course of the blood-vessels. 

The specific gravity test (p. 63) will indicate an in- 
variable increase in specific weight where infiammatorij 
exudation has occurred, no matter whether the altered con- 
sistence be great or scarcely appreciable. On the other 
hand, non-inflammatory white softening shows an invariable 
reduction in specific gravity. 

By the freezing methods it will be possible to obtain fair 
sections through inflammatory patches of greatly reduced 
consistence, and much valuable information will be thus 
obtained ; but in all cases of extreme softening all that can 
bo hoped for is obtained by raising a little of the creamy 
pulp upon the scalpel, and transferring it to a slide and 
using a thin glass cover. Sections through the membranes 
and cortex, in cases where these are involved, will jirove 
highly instructive, and should never be neglected. They 
should be studied in the fresh state, and the student become 


thorouglily familiarized with the morbid appearances ere he 
attempts to mount stained sections of such lesions for perma- 
nent preservation. Coarse blood-vessels passing through the 
inflamed patch shoidd be dra-\vn out and examined microscopi- 
cally, noting abnormal conditions of their coats and sheath. 

7. CEdematous Conditions of the Brain. — It will be 
seen from the foregoing remarks that this morbid state of 
the cerebral tissue accompanies all cases of softening of the 
brain, whether inflammatory or non-inflammatory in origin ; 
yet it is thought ad\dsable here to consider it independently 
as a condition, often the only morbid one recognized by the 
naked eye, in the brain of the chronic insane. As it is chiefly 
compensatory in its origin, it will be considered together with 
that compensatory effusion into the ventricles and beneath 
the membranes of the brain which so frequently accompanies 
it. If the cerebral tissue becomes much infiltrated with 
serum, it is liable to break down the textiu-e of the brain. 
As before stated, this may proceed to a state of white soften- 
ing, in which the brain-tissue may become quite diffluent, 
such as we observe in the neighbom'hood of the lateral ven- 
tricles in acute hydrocephalus and around inflammatory foci. 
The more frequent condition met with in the brain of the 
insane is that of a general moist condition of the white 
matter, which is soft, almost pasty, clings to the blade, and is 
apt to tear away in shreds of a dirty white hue. Such is the 
appearance in cases of senile wasting of the brain. The 
white medullary strands immediately bordering upon the 
lateral ventricles will usually be found most implicated, being 
here in closer contact Avith the serum contained in these 
cavities. The lesser degrees of oedema are recognized by a 
pecuHar and notable brilliancy of the white substance when 
cut across, and is one of the most frequent appearances in the 
brain of the insane. The student must learn to recognize the 
general moisture, swollen condition, alterations in consistence, 
and alterations in appearance from a brilliant to a dull, dirty 
white Jnie, associated with varying degrees of oedema. 


If the freezing microtome be liberally used, he cannot fail 
to appreciate to the fullest extent the varying degrees of 
oedema of the cerebral tissue ; and this not only because it 
proves the most troublesome obstacle to a free manipulation 
of delicate sections, but from the tendency of cedematous 
brain to freeze into a hard icy solid, differing much from the 
consistence of healthy brain when frozen. It is impossible to 
cut through such frozen brains except by a modified process 
to be alluded to further on. In the hardening processes, also, 
these brains undergo, by dehydi"ation, enormous shrinking. 

The serous effusions beneath the arachnoid and into the 
ventricles, which have been alluded to as often associated 
with cedematous conditions of the brain, may occur as the 
result of — 

1. Senile atrophy of the brain (compensatory). 

2. Pressure by morbid products on the vessels, as in — 

Tubercular meningitis. 
Syphilitic disease of blood-vessels. 

Growths, or abscesses of mid-lobe of cerebellum press- 
ing on venae Graleni. 

3. As a gradual accumidation from anaemia and other 
existing morbid conditions, e.g.y chronic phthisis. 

Where does the effusion of serum attending oedema of the 
brain occur? 

1. Into sac of the arachnoid, i.e., betwixt the cerebral 
layer of the arachnoid and the polished inner surface of the 
dura mater. On removal of the brain, we note this fact by 
examining the amount accumulated in the occipital fossae 
above and below the tentorium. 

2. Beneath the cerebral arachnoid. Observe how the 
latter is floated up by the subjacent fluid where this mem- 
brane bridges across the sulci. 

8. In the meshes of the pia mater. Note its swollen 
gelatinous appearance from infiltration with serum. 

4. Into the ventricles, more or less distending the lateral 


ventricles. Stretcliing tense the thin arachnoid which extends 
between the cerebellum and medulla over fourth ventricle. 

Before concluding the subject of oedematous conditions of 
brain and compensatory effusions of serum, we would recom- 
mend the student to acquire infoi'mation upon the following 
facts in all such cases : — 

1. Condition of ependyma (lining membrane of ventricles). 

a. As to healthy aspect, h. Grranular appearance 
and feel. c. Macerated aspect. cL Fibro- 
cartilaginous plates. 

2. Turbidity and coloration of serous fluid. 

3. S^^ecific gravity of serous fluid. 

4. Reaction to litmus paper. 

5. Specific gravity of the brain. 

Augmented Consistence. — All cases of indm\ation of 
the brain should induce us to examine the morbid change in 
the texture as regards its degree, its diatrihution, and nature. 
The cases most likely to present themselves to our notice 
may be embraced under the following categories : — 

1st, Greneral augmented firmness of the whole brain 

from increase of its connective or neuroglia element. 
2nd. Limited but extremely indurated patches or 

nodules due to morbid growths, especially the 

3rd. Limited indurations of grey or white matter due 

to transformation of inflammatory products. 
4th. Sclerosis disseminated or distributed down the 

motor strands. 

The student will observe that the increase in consistence 
may vary from a scarcely appreciable degree, and due 
merely to a lessened quantity of water (Rokitansky) up to 
a condition of leathery callous consistence. 

A few words upon the varieties of sclerosis met with in 
cerebro-spinal centres independent of that general increased 


firmness from increase of neuroglia observed in the brain 
of many epileptics. 

1. Superficial Scleroses. — We should also look for 
instances of partial sclerosis of the grey cortex occasionally to 
be seen in epileptics and imbeciles, and characterized by a 
peculiar shallow puckering of the surface of the gyri, 
which are somewhat indm\ated, and have a cauliflower 

2. Hypertrophy of Neuroglia. — Again we may meet 
with instances of hypertrophy of the brain, as it is termed, 
which has been found due to extreme increase of the 
neuroglia around the medullary strands of the hemispheres. 
Such a case would present us with increase in consistence, 
elasticity, and volume. In all cases of general increased 
consistence it is imperative that the greatest attention 
should be paid to examination of the brain under the 
following heads : — 

a. Volumetric measurement. 

h. Absolute weight of brain and its divisions. 

€. Specific gravity of grey and white matter. 

3. Cicatricial Formations. — The indui'ations resulting 
from inflammatory action will frequently attract attention. 
Such patches of callous tissue may surround foci of softening 
of the cortex due to plugging, or the site of old hsemor- 
rhages. Cyst-like cavities, with hard fibroid walls, may thus 
be formed deep in the substance of the hemispheres. 
Sections through such structures should be made for micro- 
scopic examination, as they illustrate well the various stages 
of transformation of inflammatory products into callous 
cicatricial tissue. 

4. Disseminated Sclerosis. — If, in slicing a brain, 
greyish nodules are found scattered irregularly through the 
white substance resembling the cineritious substance in colour. 


but becoming of a rosy hue on exposure, and exuding a 
colourless fluid, the student has probably to deal with a 
case of disseminated sclerosis. Note in such a case the 
increased consistence of the sclerosed patches, their yascu- 
larity, irregular distribution through the medullary substance, 
and the comparative immunity from this lesion enjoyed by 
the cortex. Sections should be preserved for microscopic 
examination. The extent of implication of the ganglia, 
pons, medulla, and spinal cord should also be examined. 

5. Descending Sclerosis. — If again a greyish indiu'ation 
should be met with involving the white strands running 
down from the motor region of the brain to the internal 
capsule, or from the latter down into the crm-a, and on 
into the lateral columns of the cord, we are dealing with 
a secondary descending sclerosis (fasciculated sclerosis of 
Charcot) . 


The alternating ligliter and darker shadings of the cortical 
layers vary much in the depth of their liue, rendering the 
distinctness or differentiation of the individual layers more 
or less apparent. The variations in hue from the normal 
standard may be distributed in mottled patches or laminated 
zones, or uniform ti/ spread thronrjhout the wJioJe cortical envelope. 
We should consider the conditions usually associated mth 
alterations in the coloiu- of the cortical layers, and first and 
most frequent amongst such conditions must be placed altera- 
tions in the hlood-supphj. The next most frequent condition 
met with is disintegration of nervous tissiie : next to these in 
their relative order of occurrence we find morbid deposits ; 
and lastly, new grouihs in the cortex. 

Let us first consider the morbid conditions associated -witli 
unusual pallor. 


1. Partial Mottling and Laminated Pallor. — A very 
slight acquaintance with the morbid appearances found in the 
brain of the insane will suffice to attract attention to the 
frequent presence of irregular, oval, or circular patches of 
pallor, very limited in extent, and of a yellowish grey hue. 
These patches may extend throughout the whole depth of 
the grey matter or be limited to the superficial layers alone. 
Now this condition appears almost invaiiably attributable to 
antemie zones, in which special systems of arterioles are 
involved. It may, however, be due to granular disintegra- 
tion of nerve-cells and fatty accumulations in the peri- 
vascular sheaths. 

A similar patchy pallor will also be frequently met with in 
the central ganglia, but here it will usually be found asso- 
ciated with fatty degeneration of the nuclei in the walls of 
the blood-vessels. 

A portion of a convolution exhibiting this appearance 
should be placed on the freezing microtome, with the blotchy 
aspect uppermost. On freezing it will be observed that the 
whole depth of cortex assumes a uniform pallor throughout, 
and sections cut o& and floated in water /ail to exhibit t/ie 
pale patches if the latter be due to ana3mic conditions simply ; 
on the other hand, if due to structural disintegration, the 
appearance vanishes during the frozen state, only to return 
rapidly when the section is cut. The process of hardening 
by chrome salts dissipates all patches of pallor due to anaemic 
zones, for by both processes the blood-vessels are more or less 
emptied of their contents, and a uniform tint residts. 

Apart from the presence of fatty or granular debris, the 
general pallor of the cortex may be due to great paucity of 
nerve-cells, whilst the blood-supply being diminished the 
resulting pallor will be associated with a poor differentiation 
of layers. This apparent fusion of laj'ers and loss of the 
sharp boundary lines seen in healthy brain is therefore a fact 
of some significance ; it almost invariably points to general 
malnutrition and disintegration of nerve-cells. Pallor, how- 
ever, is not necessarily associated with disintegration of 


nerve-cells, for tlie latter condition is frequently attended by 
vascular injection. 

2. General Diffused Pallor. — I cannot do tetter here 
than describe the appearances fonnd in the case of a patient 
exhausted by phthisis, in which the typical palJor and mal- 
nutrition of the nervous centres is always so prominent a 
feature. Such cases are unfortunately so frequent in oiu' 
asylums that the student will have little difficrdty in pro- 
curing such a brain for study. 

Upon removal of the extremely thin, blanched skull cap, 
and reflecting the dm-a mater, the subjacent membranes were 
seen separated "widely from the surface of the brain, buoyed 
up by fluid which in part was clear and translucent, in part 
slightly turbid from films and flakes of lymph. The small 
arterial branches were not visible, the larger primary branches 
being alone apparent at the vertex. The venous sj^stem was 
represented by engorged trunks, their minutest radicles being 
well seen. The arachnoid where it crossed the sulci was more 
or less cloudy, the milky opalescence being due to interstitial 
change and lymph deposits. In the postero-parietal regions, 
where the membranes were floated by the distending fluid off 
from the siu-face of the gyri, the smaller veins could be traced 
meandering through the fluid, and dipping down to enter the 
cortex. Tracing the course taken by these vessels, the smallest 
radicles were seen emerging from the cortex, uniting here 
and there with larger twigs, which ramify through the serous 
fluid and cross obliquely or directly over the summit of the 
convolution, to terminate in the primary veins which are 
formed by their convergence. These latter veins run up the 
sulci on either side of the convolution, superficially exposed 
and directed towards the median line or longitudinal fissm'e, 
where they terminate in the longitudinal sinus. To render 
their distribution more apparent, we have only to compress 
the orifice of one of the larger superficial veins near tlie 
median line, whilst Avith the handle of the scalpel placed on a 
distal part of the same vein, we include numerous converging 


radicles betwixt the occluded points. Upon gradually draw- 
ing the handle of the scalpel along the course of the vein 
upwards, the blood is projected into these radicles, and innu- 
merable branchlets before invisible strike across the field over 
the convolution, or meander in tortuous arborescent forms 
through the subjacent fluid. The student will learn from the 
above examination the following relevant and useful facts : — 
1st. The larger arteries being at the base, the smaller 
branches are alone seen near the vertex, and 
these usually deeply seated in the sulci. 
2nd. All the larger blood-vessels usually exposed at the 

vertex are veins. 
3rd. The direction taken by the minute venous radicles 

will be accurately learnt. 
4th. A high degree of vascularitj^ simulating passive con- 
gestion may be induced artificiaUy with ease by 
slight force. 

It is necessary to observe here that great caution should be 
taken by the student against hastily arriving at any conclusion 
as to the presence of congestion of the brain from a super- 
ficial view of the veins at the vertex — upon this point the 
tyro constantly errs. " We must make it a rule to consider 
hypercemia of the cerebral membranes as proved only in those 
cases where the finest vessels are also injected, and where the 
overloading of the cerebral vessels is not at all in proportion 
to the amount of blood in other organs." ^ So says Niemeyer, 
reiterating a caution constantly given by pathologists, and as 
frequently neglected. Proceeding with our study of the 
anaemic brain, the membranes were next stripped and two 
facts noted ; they were slightlj^ thicker and more tough than 
in health, and ^Qj Avere removed with extreme ease. The 
student must be familiar with the difiicult}^ experienced in 
stripping the membranes from healthy brain, being recom- 
mended often to remove them under water. This difficulty 
is not alone due to the tenuity of the pia mater and arach- 

1 Niemeyer' s " Practical Medicine," translated by Drs. Humphrey and 
Hackley, vol. ii. p. loo. 


noid, but also to the fact that the former membrane is attached 
to the siu-face by the prolongations of connective cells recog- 
nized as Deiter's corpuscles. These are by no means nume- 
rous in health, yet sufficiently so as to keep the pia mater in 
firm contact with the cortex. These cells are chiefly found 
in the neighbourhood of the vessels, where they enter the 
cortex from the pia mater, and both together they form an 
impediment to easy stripping. In cases of serous subarach- 
noid effusion, such as the one under consideration, the mem- 
branes have been floated up from the receding surface of the 
atrophic cortex, and the connections with Deiter's corpuscles 
have therefore been forced asunder. After noting fully the 
consistence of the brain accordmg to the plan ah-eady given, 
a section was made exposing the centrum semi-ovale, and the 
following facts presented themselves : — 

1st. The blanched aspect of the cortex and the absence of 
any reddish striation of the upper layers, such as is seen when 
the cortical vessels are fidl of blood. The hue of the cortex 
was asht/ grey throughout, except in its most vascular zone, 
where perhaps the slightest warmth of tint was recognizable, 
but beyond this, nowhere did any blush suggest the presence 
of its extremely elaborate vascular apparatus. 

2nd. The layers were individually very poorly differen- 

3rd. The medulla was brilliant white, glistening, slightly 
reduced in consistence, and showed few or no puncta vas- 

"We must bear in mind that the vessels of the meninges 
may be tortuous and tm-gid with blood whilst the minute 
vascular supply of the cortex is absolutely diminished, and 
the layers appear blanched and anoemic. To a certain extent 
this was the case in the brain we have under consideration. 
How this occurs (passive or active hyperremia of the mem- 
branes associated with anaemia of the cortex) will be explained 
in our next section when speaking of hyperoemia of the brain. 
For the present let us mark well the fact that the vascular 
condition of the membranes is no index, as a rule, to the state 
of the cerebral substance. 


The brilliant white glistening aspect of the medulla was 
due, as will be shown fmiher on, to a slight degree of oedema 
of the brain-substance. 

Let us now place a small portion of a parietal convolution of 
thin and anaemic brain upon our freezing microtome, and cut 
a fine section through the cortex — float it on the glass slide 
and drain off superfluous fluid. Take a hand lens and 
examine first by reflected light. 

The cortex is seen as apparently to consist of — 

1st. An outer light translucent zone extending half way 

2nd. A. deep opaque white layer extending to the 

medulla, which is still more opaque and white 

in aspect. 

Eaise the slide and examine by transmitted light. We 
now see — 

1st. The narrow translucent grey of the first or outer 

layer of the cortex. 
2nd. A broad less translucent grey zone of the second 

and third layers. 
3rd. A broad stripe of darker grey extending to the 
medulla, but separated at one-third its depth 
by a narrow bluish belt. 

We see thus by the naked eye the constitution of the 
cortex in this region of the brain, except that the narrow 
second layer cannot be defined from the third layer by 
unaided vision. Let us tiirn to the cortex prior to section 
cutting and examine by means of a hand lens imder re- 
flected light, and the appearances are as follows: — 

1st. A grey belt corresponding to the first layer. 
2nd. A broad white belt divided midway by a narrow 

grey line. 
3rd. A grey belt constituting the deepest layer. 

We have, I think, gone sufficiently into the appearances 
afforded by the ansemie brain for our present pui-pose. The 


examination of the various layers above recommended will 
prepare us not only for recognizing the relative positions of 
layers which have later on to be minutely studied, but will 
teach us to estimate roughly — 

a. The differentiation of the layers by naked vision. 
h. The relative depth of individual layers. 

c. The atrophy or normal depth of the cortex. 

d. The presence of morbid products. 

Of the latter I need here only mention those cases of so- 
called miliary sclerosis in which the nodules can be distinctly 
seen in sections on examination by reflected ligJtt, dotting the 
surface over mth minute opalescent spots. Let us now 
recapitulate shortly the observations to be made when 
examining by naked vision a cortex of unusual pallor : — 

1st, The extent of pallor throughout the brain. 
2nd. Its limitation in depth through the cortex. 
3rd. Its disposition in patchy areas or otherwise. 
4th. Its association with obscure laminar boundaries. 
5th, Its association mth pigmentary tints. 
6th. Its association with altered consistence and oedema. 
7th. Its reappearance in sections from frozen brain. 
8th. Examination of sections by transmitted and reflected 

Redness. — The depth of tint acquired by the grey layers 
of the cortex depends not alone on the presence of pigmented 
nerve-cells, but also on the far greater vascularity of the grey 
as compared with the white matter, whilst the layers most 
richly supplied with nerve-elements possess also the more 
abundant capillary supply. The more vascular the layer is, 
the deeper and warmer will be its tint. Now the capillaries 
of the cortex are of remarkably fine calibre, and hence, when 
the ultimate arterioles are injected, the appearance resulting 
is that of a uniform rosij hlunh more or less dark, the surface 
smooth and sicollen. This uniform coloration, if at all extreme, 
is often attended by slight extravasations of blood, and the 

E 2 


tissue around presents a different staining from blood-pigment. 
Such are the conditions found in genuine congestion of the. 
cerebral substance, and the analogous condition is readily 
recognizable in the soft membranes. The student must make 
himself thoroughly acquainted with the evidences of a 
genuine active or passive congestion of the brain, and learn to . 
distinguish it from actual inflammation, where, as ak-eady 
pointed out, the consistence as well as colour are profoundly 
implicated, and the specific gravity also affected. 

1. Congested Zones. — One of the most frequent appear- 
ances in the cortex of the acute forms of insanity, is a bright 
arterial zone, which bounds the confines of the white and 
grey matter of the convolutions, following out accurately the 
direction of the innermost cortical layer. This congested 
belt corresponds to the horizontally disposed nexus of 
blood-vessels into which the larger straight cortical arteries 
empty themselves after passing through the various cortical 
layers. Any undue engorgement of the vessels of the pia 
mater will necessarily affect these larger blood-vessels, and, 
from their general arrangement, a mechanical element is 
brought to bear, such that, as I shall endeavour to show 
fiu-ther, is significant of a safety-valve action for the cortex 
in cases of vascular engorgement, relieving the cortex some- 
what by their distension from congestive conditions. It is 
on this account, I believe, that this linear vascularity is so 
often apparent without a corresponding blush in the cortical 
layers, indicative of distension of the minute capillaries. 

2. Limited Patchy Congestion of Cortex. — When, 
however, the cortex is itself congested, the student will 
frequently observe irregular bright red patches, suggestive, 
like the anaemic patches already referred to, of the implication 
of minute vascular areas, whilst leading down to them are 
seen the distended straight cortical vessels, giving the upper 
layers a reddish streaked aspect. The last act of arterial 
contraction, in ^hich the smaller arterioles have failed to 


empty themselves into the venous system, may in part explain 
this appearance, and caution is necessary, lest we hastily 
assume that this state is the result of morbid activity. The 
student should, therefore, examine closely the injected part 
by the microscope, and look for evidence of minute extrava- 
sations, hsematine crystals, and staining as well as broken- 
down texture, or other results of inflammatory action and of 
congestion. This blotchy red aspect of the cortex reappears 
very frequently in the medulla in similar cases, and is indica- 
tive of engorgement of the minute capillaries of these regions. 

3. Puncta Vasculosa. — It is customary, in judging of the 
degree of engorgement of the vessels of the white matter, to 
be guided by the paucity or abundance of the puncta vascu- 
losa, caused by section of the engorged channels — in other 
words, the number of drops of blood oozing from the 
divided vessels are supposed to form some criterion of the 
engorged condition of the vascular system here. It has been 
shown by Niemeyer that this is no safe criterion, as 

1st. The number of puncta vasculosa vary greatly with the 
fluidity of the blood, which is an important element in their 

2nd. They are often almost entirely absent when we have 
had indisputable evidence of intense vascular engorgement 
during life. 

We must, therefore, be on our guard against attributing too 
much importance to abundant bloody points on section of the 
medulla, whilst we should never fail to note whether this 
condition is or is not associated with distension of the minute 
capillaries as indicated by fine diffuse patchy redness. 

I need not here dwell upon the changes of coloiu' due to 
inflammatory action, as these have been already disposed of 
when illustrating anomalies of consistence ; but I must here 
repeat the important caution that the student be not misled 
by expecting to find engorgement and redness in all cases of 
encephalitis. The larger number of such cases present no 
undue red coloration. 


Results of Intra-cranial Pressure, — Due allowance 
must be made for conditions of increased intra-cranial {hut 
extra-vascular) pressure. Wlien the cranial cavity is en- 
croached upon by a tumour, abscess, hsemorrhage, or serous 
accumulation, compensation is made by the outflow of the 
subarachnoid fluid into the spinal meninges. The limit of this 
compensatory arrangement is soon reached, and then the only 
available space is that gained at the expense of the general 
vascular calibre. Encroachments upon the cranial cavity 
therefore eventually empty the blood-vessels, causing partial 
or more or less general anaemia prior to compression of the 
brain-substance itself. It is therefore far from improbable 
that, as Niemeyer teaches, engorgement of the blood-vessels 
may reach such a degree, that after the limit of subarachnoid 
compensation has been reached, effusion of serum occui-s into 
the perivascidar spaces and brain-tissue, and suiflces to com- 
press the minuter blood-vessels and capillaries. Hence we 
may find in such cases an association of marked cerebral 
anaemia with tortuous and engorged blood-vessels in the 

Results of Intra-thoracic Pressure. — Venous engorge- 
ment of the brain is a fi'equent result of obstructed circula- 
tion through the medium of pleuritic effusions, intra-thoracic 
growths, fibroid ind oration, and other changes obstructing or 
obliterating the vascular system of the lungs. Occasionally, 
although rarely, there is found an extraordinary engorgement 
and varicosity of the veins of the pia mater — the vessels 
winding in all directions, and, as Eokitansky states,i even in 
spirally -twisted coils and intestine-like ciiTiunvolutions. In 
these cases the brain-substance will be found dark and 
engorged, full of puncta vasculosa and even miliary apo- 
plexies. In one very typical case occurring at West Riding 
Asyliun the brain-surface was actually concealed over exten- 
sive tracts by a vast development of varices and contorted 
vessels, and when cut into, the grey and white substance was 

' " Patliological Anatomy," Sydenham Soc. vol. iv. pp. 372-3. 


not only deeply engorged, but presented very numerous 
miliary haemorrhages, forming patches varying from a pea to a 
florin in extent, and in all degrees from the purely punctif orm 
to the diffuse uniform extravasation. A similar case was ex- 
hibited some years back by Dr. Coupland at the Pathological 
Society of London.^ In this case the mechanical hyperaemia 
was due to acute bronchitis — in the Asylum case the imme- 
diate cause was the supervention of capillary bronchitis upon 
fibroid limgs in a patient who gave a history of intemperance. 
Its association with diink has been recognized by Rokitansky. 
The occm-rence then of such cases should direct the student's 
attention to the effects of intra-thoracic pressure upon the 
venous circulation of the brain. 


Volumetric Methods for the Brain. — The cubic 
measm^ement or volume of the encephalon can be very readily 
ascertained, and considering the important information it 
affords, it is a process too frequently neglected in our post- 
mortem rooms. The volume of the brain is estimated by its 
displacement of fluid. 

1. Dr. Hack Tuke has detailed, in the January number 
of the British and Foreign Medical Review for 1855, the 
result of examination by this method of sixty-three brains, 
together with the capacity of the crania.. The vessel used by 
this observer was one of convenient size and shape, with a 
capacious spout placed at an acute angle with the sides. 
Water is poured into this vessel up to the level of the spout. 
Fluid contained mthin the ventricles and subarachnoid space 
is allowed first to escape by several long incisions, and then 
the brain, including the medidla oblongata, is immersed, 
the displaced water, as it escapes from the spout, being caught 
and measured, affording an exact criterion of the actual bulk 
of the brain. The student may, however, prefer the use of a 

' Reported in the Lancet for January 11, 1879. 


graduated vessel upon which lie can at once read off the dis- 
placement of fluid, but the exact graduation of large vessels, 
such as would be required, is open to several fallacies ; and 
these possible sources of error are, of course, wholly avoided 
by Dr. Hack Tuke's method. If such a vessel as that used 
by Dr. Tuke be not at hand, a ready method is the one I 
have used, as follows : — 

2. Half fill with water an inverted bell-glass of sufficient 
dimensions, and mark the level of the fluid. Now immerse 
the brain in the fluid, and note the level of displacement, 
after which the brain is removed ; and, if the original level 
is not exactly maintained owing to imbibition by the brain, 
pour in sufficient fluid to compensate for the loss. Water is 
now poured in from a graduated measure up to the displace- 
ment level, the amount required for this pui^pose giving us the 
volume of the brain. The volume should be estimated in 
cubic centimeters and cubic inches. 

I employ for section-cutting a vessel which answers well 
for the volumetric estimate of the brain. It consists simply 
of a large glass vessel, such as is used for the preservation of 
brain in museums, but fitted with a stopcock arrangement 
in the side near the bottom of the vessel. It is filled with 
sufficient water to cover the brain, the level being marked or 
indicated by a weighted float. The brain is next immersed, 
and whilst so immersed, the fluid of displacement is run 
off by the stopcock, and measm-ed in a graduated vessel. 
Again, in lieu of a graduated vessel, the student may emj^loy 
a float, movable along a graduated weighted stem (gradu- 
ated, of coiu-se, for the vessel used). Such a float should 
terminate below in a brass button, which rests against the 
bottom of the vessel, keeping the graduated stem in a 
vertical position, whilst the float freely moves with the 
rising or falling level of the fluid along the graduated stem. 

3. When referring to the specific gravity of the brain 
an apparatus will be described whereby volume as well as 
weight of brain are readily measured.^ Such an apparatus 

^ Stevenson's Displacement Apparatus, vide p. 65. 


has the recommendation of being cheap and efficient. 
Whatever arrangement be adopted we should not rest 
content with the estimate of the volume of the encephalon 
alone. Each hemisphere should be separately measured, as 
also . the cerebellmn, pons, and medulla. It will be found 
convenient to use smaller and more delicately-graduated 
vessels for the latter. 

Cranial Capacity. — This should be estimated Avith a 
■vdew to a comparison between the actual volume and weight 
of the brain and the dimensions of the cranial cavity, as 
well as for comparison with the capacity of average-sized 
skulls. The relationship between cranial capacity and brain- 
weight has been sho\\Ti by Dr. Barnard Davis, who has 
adopted the general rule that a deduction of about fifteen 
per cent, from the capacity of the cranium gives the 
^' capacity " of the brain, whereby its weight may be readily 
calculated.^ The large bulk of cranio-metric observations 
having been taken upon the cliied skull, we should, when 
dealing with the fresh subject, make allowances for slight 
divergence ; and, in order to approximate the conditions, 
the dm-a mater should be wholly removed from the skull- 
cap and the base. When, however, our intention is to 
estimate the difference in any single case between cranial 
capacity and brain-volimie, rather than for more general 
comparative purposes, it will be necessary to open the skull 
prior to opening the thorax, so as to avoid emptying the 
venous sinuses, whilst at the same time the diu-a mater must 
not be stripped away from the skull-cap nor the base be 

1. With the object of estimating the amomit of cerebral 
atrophy, a valuable series of observations were made by 
Dr. Hack Tuke, and the process he adopted is described as 
follows : — " The foramina at the base of the brain are 

^ *' On the Weight of the Brain in the different Races of Man." Thilos. 
Trans., 1868, pp. 506 and 526. 


carefully plugged witli tenacious clay — that used by statu- 
aries for modelling answers best ; a small triangular piece 
of the frontal bone is removed with the saw ; the calvarimn 
is readjusted to the base, the dm^a mater being left attached. 
The space left by the attrition of the saw in removing the 
calvarium is filled with clay ; and a narrow bandage with 
clay spread upon it is made to surromid the cranium three 
or four tunes, covering this space. If this manipulation 
has been carefully done, the cavity of the cranium will now 
be found as tight as a bottle. Sixty fluid ounces of water 
having been measured, a sufficient quantity to fill the cranial 
cavity is now poured into it by means of a funnel, through 
the orifice in the frontal bone, taking care that the stream 
does not wash away the luting of the foramina. The fluid 
which remains, after having filled the cranial cavity, is 
measm^ed, and being deducted from the sixty ounces gives 
the amount emj)loyed. To this must be added half an ounce 
for the space occupied by the luting." Having thus obtained 
the cranial capacity, he deducts from it the brain volimie, 
and obtains thus the exact measui^ement of shrinking or 
atrophy. Millet seed and sand have been used for measur- 
ing the capacity of the cranial ca^dty, and the latter in the 
dried skull has answered admirably. It must, however, be 
borne in mind that a fallacy may be introduced by the 
employment for this purpose of materials liable to be 
influenced by temperatm'e. Fluids are, of course, more 
open to this objection, their expansion by heat being pro- 
portionately more rapid than solids, and hence it would 
appear that sand is preferable to water in these investiga- 

2. Mustard seed was employed by Professor Flowers in 
his extensive series of researches.^ 

3. The method adopted by myself arose from my em- 
ployment of solid paraffin in obtaining casts of the brain- 

' The capacity of the crania contained in the Himterian Museum of the 
Royal College of Surgeons was obtained by Professor Flowers in this 


surface, and the interior of tlie skuU. It was soon apparent 
that the finest and most delicate impressions and most perfect 
casts coidd be obtained by the use of this substance, and that 
it did not share in the great disadvantage which accrues 
from the use of plaster of Paris, viz., that of great con- 
traction during solidification. It is in this respect also far 
superior to white wax, which, as is well known, contracts 
much whilst solidif jing. I proceed as follows : — Fill up 
the foramina at the base as pre\iously described. A tri- 
angular or wedge-shaped piece is now sa^vn out of the 
occipital bone after removal of the calvarimn, but retained 
in situ. The base is then filled up by melted paraffin, the 
skrdl-cap replaced and fastened by luting, just as in Dr. 
Tuke's process, having previously trephined a piece out of 
the frontal bone. Through the latter orifice more of the 
paraffin is poiu"ed in imtil the cranial cavity is filled. When 
cool and solid remove the calvarium, as well as the wedge- 
shaped piece of bone from the occiput, and then gentle 
pressure from behind tilts the solid mass out of the cranium, 
Avhen it will be found to form an exquisite mould of the 
interior. The moidd thus obtained is now to be measured 
by displacement, whence we obtain the cranial capacity. 
This method has afforded me great satisfaction, as it gives 
data of great value at the expense of very little trouble : 
it supplies us with a method of — 

1st. Estimating cranial capacity. 
2nd. Grives us an exact mould of the cranial cavity. 
3rd. As a permanent record — numerous linear angular 
measurements and volume measiu-ements may be obtained. 

4. From some invaluable measurements by the late 
lamented Broca,^ it was found that of 115 skulls of indivi- 
duals living in the tweKth century, the average capacity was 
1425 "98 cubic centimeters ; whilst of 125 skulls of the nine- 
teenth century the average capacity was 1461'53 cubic centi- 
meters. The following table of percentage will be found of 

1 Quoted by Vogt, " Lectures on Man." Anthrop. Soc. 



use by those interested in the subject of cranial capacity. It 
is given by Le Bon as illustrative of the relationship of race 
to cranial capacity : — 

Cranial Capacity in different Human Races. 





Parisians of 
12th Cent. 




Cubic Centimeters. 

1,200 to 1,300 . 





1,300 to 1,400 . 






1,400 to 1,500 . 






1,500 to 1,600 . 






1,600 to 1,700 . 






1,700 to 1,800 . 






1,800 to 1,900 . 







Gravimetric Methods. — The student must make him- 
self familiar with the various circumstances which modify 
brain-weights. Some of these conditions have been sum- 
marised by Bastian.^ They appear to be chiefly as follows : — 

1. Length and natm^e of illness. 

2. Mode of death (vascidar engorgement favom'ing high 

3. Certain neuroses, as epilepsy. 

4. All conditions inducing sclerosis. 

5. Atrophy. 

6. Congenital micro- or megalo-cephaly. 

The student must also take into consideration the relationship 
between weight of brain and the age and sex of the individual 
as well as the weight and height of body. 

^ ''The Bruin as an Organ of Mind," chap. xx. 


Absolute Weight of Brain. — Before weighing tlie 
brain, all fluid from the subarachnoidal space and ventricles 
must be di-ained off, and allowance made for the membranes, 
which will scarcely exceed an oimce in weight. The plan 
adopted at the "West Riding Asylum, however, is to first 
strip the brain of its membranes, to liberate the fluids in the 
ventricles, and weigh the whole encephalon. The membranes 
can afterwards be weighed if thought necessary. In all 
cases when the pia mater is firmly adherent to the cortex, it 
is as well to weigh the brain prior to stripping, as large 
shi'eds of the cortex are often removed in these cases. Pre- 
caution must be taken, however, by incisions, to release any 
accumulation of fiuid in the subarachnoid spaces and meshes 
of the membrane, and, subsequent to weighing, allowance 
must be made for the pia mater and arachnoid. After the 
weight of the encephalon has been obtained, the cerebellum 
and pons must be removed by di\iding the crura cerebri close 
to the pons} A longitudinal incision is then carried through 
the median line of the corpus callosum from before back- 
wards, so as to separate the two hemispheres. The pedun- 
cular connections of the pons and medulla with the cere- 
bellum are then divided, and the former (pons) also separated 
from the medulla at its natiu-al line of division. This is the 
method adopted at West Riding Asjdum, but for several 
years it was the custom here to separate the frontal lobes 
from the remaining posterior part of the brain by an incision 
carried through the fissure of Rolando, The weights of 
these individual parts are then taken. 

It will be foimd ad\dsable to use the metric system in aU 
our estimates of capacity, volume, and weight, and all the 
standard brain-weights afforded by the elaborate tables of 
Tiedemann, Reid, Boyd, Wagner, and others shoidd be 
expressed in grammes rather than ounces. Dr. Sharpey, 
after an elaborate analysis of brain- weights given by Grlen- 

' A modification of the plan is recommended in tlie section on the dis- 
section of the brain as preferable when the question of weight is of secondary 
import (p. C9). 


dinning, Sims, Tiedemann, and Reid, supplies us with the 
following valuable results : — ^ 



weight of adult Male Brain 



» )) 5) 



») )) )» 



„ „ Female Brain 



» » j> 



)) 5> »> 


The heaviest human brain on record has been described by 
Dr. Morris,^ and was carefully examined at University 
College Hospital : the weight was 67 oz. 

Compatible with ordinary intelligence, the lowest limit of 
the human brain, as regards weight, is, according to Gratiolet, 
900 grammes, and, according to Broca, 907 for the female, 
and 1049 grammes for the male.^ 

Specific Gravity. — Researches upon the specific gravity 
of the encephalon are becoming more and more interesting 
with our knowledge of cerebral localization and intimate brain- 
structure. The late suggestive work of Danilewsky upon the 
relative amount of grey and white matter in the brain depends 
greatly upon the accuracy of the specific gravity for its cor- 
rectness. The more important methods adopted are those of 
Drs. Bucknill, Sankey, and Peacock. 

1 "Elements of Anatomy," 7th Edition, vol. ii. p. 568. 

2 Brit. 3Ied. Journ. Oct. 26, 1872, p. 465. 

2 Quoted by Bastian, "The Brain as an Organ of Mind," 1880, p. 365. 
In connection with the weight of the brain in the insane, the student 
will find most valuable information in articles by Dr. Crochley Clapham, 
contained in the 3rd and 6th vols, of the "West Riding Asylum Medical 
Reports." His observations embrace 1,200 cases of insanity. Fiu-ther 
information upon this subject may be obtained by reference to the 
following : — Sims, Medico-Chirurt/. Trans, vol. xix. ; Glendinning, Medico- 
Chirurg. Trans, vol. xxi. ; Tiedemann, " Das Hirn des Negers," Heidelberg, 
1837 ; Reid, London and Edin. Month. Journ. Med. Science, April, 1843 ; 
Thurman, Journ. Mental Science, 1866 ; Wagner, " Vor.studien," 1862, 2' Abh. 
pp. 93-95 ; Peacock, Month. Journ. Med. Science, 1847, and Journ. of Tathol. 
Soc. 1860; Boyd, Philos. Trans. 1860; Bastian, " The Brain as an Organ 
of Mind," 1880 ; Clapham, loc. cit. and Journ. of the Anthropolog. Inst. 
vol. vii. p. 90. 


1. Buckniil's Method. — Dr. Biicknill was tlie first who 
originated a satisfactory and ready method for estimating the 
specific weight of brain. The foUomng is his detailed account 
of the process : — " The specific gravity of the cerebrum and 
cerebellum is ascertained by immersing a portion of each in a 
jar of water wherein a sufiicient quantity of sulphate of mag- 
nesia has been dissolved to raise the density of the fluid to the 
point required, adding water or a strong 
solution of the salt, until the cerebral 
mass hangs suspended in the fluid 
without any tendency to float or sink, 
and then, by testing with the hydro- 
meter, the specific gravity is thus f oimd 
with great delicacy and facility, a 
difference of half a degree in the density 
of the fluid being indicated by the rise 
or fall of the substance immersed. The 
soluble salt is chosen, for its possessing 
no astringent or condensing action upon 
animal tissues."^ 

2. Sankey's Method. — This pro- 
cess is the one which has been used 
most extensively by Dr. Crichton 
Browne at West Riding Asylimi, and 
it is the method which I have myself 
invariably adopted. It appears to me 
in every respect highly satisfactory and 
simple. "We require a series of cylin- 
drical glass jars, such as the one figured, 
and a set of graduated hollow glass bulbs, which can 
be readily obtained.- 

These glass bulbs are accm-ately graduated, or r 

Fig. 2. 
Specific Geavxty 


marked with the specific gravity of the fluid in which they 

1 Lancet, 1852, vol ii. p. 589. 

2 Such a set of graduated bulbs may be obtained of Mr. Stevenson, 
Philosophical Instrument Maker, No. 9, Forrest Road, Edinburgh. 














would liang suspended when immersed, neither tending to 
float nor sink. The glass jars are partly filled with water, 
and then a concentrated solution of Epsom salts added 
to each, until its specific gravity is such that whilst one 
bulb floats, a bulb two degrees higher sinks. Thus a series 
of jars are filled as represented below : — 

Bulbs floating .... 1030 
Specific gravity of fliiid = 1031 
Bulbs sunk 1032 

The middle line represents the specific gravity of the fluid, 
and therefore of the portion of brain which tends neither to 
float nor sink, but to remain suspended wherever placed. It 
will occur to the student that the jars may be so graduated, 
that whilst one bulb floats, another, one degree higher, sinks, 
and hence that half a degree specific gravity may be indicated 
as follows : — 

Bulb floats 1030 1031 

Specific gravity of fluid and suspended body 1030'5 1031-5 
Bulb sinks 1031 1032 

This is a degree of nicety, however, to which the use of the 
beads shotdd not be pressed, and if required, should demand 
in preference the use of the 1000-gramme specific gravity 
bottle. A series of jars should be graduated from 1028 to 
1050, so as to enable us to deal withwhite or grey matter of 
cerebrum, cerebellum, and central ganglia. Occasionally still 
more dilute solutions mil be requisite. 

A minute piece of white or grey matter is now raised by a 
scalpel and placed upon the perforated spoon or scoop, which 
is gently lowered into one of the jars of saline solutions, and 
the fragment of brain tm-ned off the scoop and closely 
observed. If it sinks, it is of course of higher specific gravity 
than the fluid, and must be passed on to stronger solutions 
until, reaching one of its o^ti density, the fragment remains 
stationary where placed. If, on the other hand, it floats, it 
must, of coiu-se, be moved to a solution of less specific weight, 
until the same conditions are obtained. Should the specific 


gravity beads be cbosen by the student for tliis purpose, it 
would be well for him to attend to the following rules : — 

a. After obtaining the specific gravity of any one portion 
of brain, repeat the trial with fresh portions at least two or 
three times, so as to ensure perfect accuracy. 

b. Examine the brain as soon as possible after death, 
wholly rejecting such as show the least evidence of com- 
mencing decomposition. 

c. Keep the jars covered with glass squares in a cool room, 
so as to avoid dust and evaporation as far as possible. 

d. Before commencing any series of observations, note the 
position of the bulbs, any tendency of the lower bulbs to float 
from evaporation of the solution being rectified by the addition 
of water. 

3. Stevenson's Displacement Apparatus. — A simple 
and most reliable apparatus has been devised hy Mr. Stevon- 

FiG. 3. — Stevenson's Displacement Api-aeatus. 

son for estimating at the same time both the bulk and specific 
gravit}' of large irregular bodies by the principle of displace- 



ment. Tlie apparatus consists of a jar, A, fitted -witli a 
large drooping tubidatiu'G, B, with stopcock attached. A 
metal gauge, D, having a thin horizontal straight edge inside 
the jar, determines the exact water-level, and over it the 
surplus water flows. The other jar, E, is graduated on the 
sides into cubic inches, avoirdupois lbs. and ozs., and thousandth 
parts of a gallon. The packing case, Gr, serves as a stand for 
the jar. In taking the specific gravity of the brain, or any 
body a little heavier than water, the jar must be filled with a 
solution of salt or sulphate of magnesia up to the level of 
the gauge, the stopcock being closed. The brain is then 
immersed in the fluid, in which it ought to float, and the fluid 
immediately rises in the jar; let it come to rest; open the 
stopcock, and the quantity displaced Avill flow into the 
graduated jar and represent the u-eight-indication of the brain 
(since any floating body displaces exactly its own weight of 
the fluid) . The stopcock is now shut, and the brain depressed 
by the brass piece, F ; and when the water has again come 
to rest, open the stopcock and receive the f m-ther displacement 
in the jar E. The whole bulk of displaced fluid read off the 
graduated jar indicates, of course, the volume of the brain. 
To obtain the specific gravity, multiply the weight-indication 
by the specific gravity of the fluid and divide by the vokmie- 
indication. I need scarcely indicate to the student the 
simplicity and value of this method, whereby the weighty the 
volume, and the specific gravity of the brain may be simul- 
taneously obtained.^ The apparatus, as figiu-ed, is somewhat 
smaller than what would be required for brain. 

In estimating the specific gravity of the encephalon, the 
student will soon appreciate the fact that the specific weight 
varies not only in the cerebrum, central ganglia, and cere- 
bellum, but that variations occur over different regions of 
the cerebrimi, suggesting the importance of a comparative 
examination by this method of the various cerebral con- 
volutions. By cautious manipulation he will also be able to 
show that the specific gravity of the cortex varies with its 

^ This apparatus is sold by Mi\ Stevenson, 9, Forrest Road, Edinburgh. 


deptli. Dr. Sankey otserves that the specific gravity dimin- 
ishes in the ratio of •001 for every twenty-four hoiu's after 
death. This must, however, be greatly modified by the 
temperatiu^e of the surrounding atmosphere, and other condi- 
tions favouring putrefactive changes. The folloAving embrace 
the more important residts obtained by different observers, and 
they will serve as a useful guide to the student in his prosecu- 
tion of similar observations ^ : — 

Average specific 
















Grey matter 




White matter 


(Sankey) . 


Central ganglia 



Proportion of White and Grey Matter in the Brain. 

— Tlie specific gravity of the grey and white matter, 
together with that of the whole brain, has been applied to 
solve the very important problem of the relative percentage 
of grey and white matter in the human brain. Danilewsky 
has lately published his results and method of procediu'e, an 
account of which may be seen in the Ccntralblatt f. d. Med. 
Wissenchaftci), No. 14, 1880. ^ The formula given by him is 
as follows : — 

X = — Y- ^ where x is the quantity of grey or of white 

matter, p the specific, grcmty of the whole brain, a of the grey 
and h of the white substance, and P is the weight of the 
Avhole brain. He gives, as the results of one series of experi- 
ments, a percentage for the grey matter of 37'7 to 39, against 
a percentage of 60'3 up to 61 for the white substance. By 
taking the average depth of grey matter it is, of course, an 

^ References to work done in the Specific Gravity of the Brain : — Bucknill, 
Lancet, vol. ii. 1852, and the Med.-Chir. Rev. 18-55; also " Psj^chologica 1 
Medicine," Bucknill and Tuke, 3rd edit. pp. 520 and 587; Sankey, Brit, 
and For. Mcd.-Chir. Rev. 1853 ; Peacock, " Trans. Pathol. Soc. of London," 
1861-2 ; Aitkin, " Science and Practice of Medicine," 1865, vol. i. p. 265. 

^ Vide Abstract by Dr. Geoghegan, Journ. Mental Science, p. 437, Oct. 

T^ 9 


easy matter to obtain tlie surface measurement of the brain. 
Wagner's researches upon the superficial area of the brain are 
of interest here.^ The student, however, must be warned 
against regarding results so obtained as other than merely 
approximate^ as it is next to impossible to exclude the 
numerous fallacies to which the estimation of average specific 
gravity is liable. Thus the specific gravity of the grey 
matter is known to vary with its depth, with its local distribu- 
tion or area, with the amount of vascularity of the tissue, and 
time after death. Beyond this the very cases in which the 
pathologist would be interested in estimating the relative 
proportions of grey and white matter are those which are 
subject to such diffuse and local change in consistence that 
an average specific gravity for either white or grey could not 
be attained with any degree of exactness. For comparative 
investigation of healthy brain, the process adopted by 
Danilewsky promises to afford valuable results as long as 
the greatest care is observed to exclude fallacies. 

1 Quoted in Quain's " Anatomy," vol. ii. 




It may be usefiJ here to review the course to be piirsiied in 
removing and preparing the various parts of the brain for 
coarse examination, and the following method is recommended 
as in every way the more reliable and satisfactory one. 

In the fii'st place the head should be so supported as to 
command a good light and a full and satisfactory view of 
the brain i)i dtii. 

In the next place never neglect the rule of o]iening the 
skull and examining the brain ^j/vor to opening the thorax 
and dividing the great vessels. 

Cranial Membranes and External Aspect of Brain. 
— The skull-cap having been removed sec. art. we proceed as 
follows : — 

' 1. Examine upper surface of dura mater in its relation- 
ships to the skull (pp. 3-5). 

2. With a forceps raise a fold of dm-a mater anteriorly, 
run the scalpel through it, and carry the blade along either 
side on a level with the sawn edge of the cranium far back 
towards the occiput, stopping short of the middle line. 

3. Reflect these lateral halves of the membrane towards 
the middle line, observing the condition of the large super- 
ficial veins running into the longitudinal sinus. 

4. Replace the dura mater, and with a curved bistomy 


open up the longitudinal sinus in • its whole length back 
towards the torcular herophili, noting the condition of the 
parts (p. 5). 

5. Divide the connection between dura mater and crista 
galli, and seizing the anterior end of the falx cerebri, 
forcibly draw the membrane backwards, dividing the 
junction of the superficial veins in this course, and thus 
expose the surface of the hemispheres. 

6. With a good light thrown on the subject, and, if 
requisite, the aid of a hand lens, observe the appearances 
presented at the vertex, noting the condition of the vessels, 
membranes, and general conformation of the brain. It is 
absolutely necessary to pay attention to this point, as the 
appearances presented in situ are often greatly modified or 
wholly lost upon removing the brain at a later stage. 

7. Grently raise the tips of tlie frontal lobes from the 
orbital plates, carefully removing the olfactory bidbs with 
the brain, and then using a little gentle traction, the optic 
nerves are exposed and divided close to their foramina. 
With the same blunt-pointed curved bistomy divide suc- 
cessively the infundibulum, carotid arteries, and third nerve. 
A good view is thus afforded of the tentorium, which should 
be divided along its attachment to the ridge of the temporal 
bone, dividing at the same time the fom-th nerve. The base 
of the brain is now exj^osed, and the fifth, sixth, and seventh 
nerves readily divided, after which the blade is passed do-wn 
into the vertebral canal on either side of the medulla, and, 
cutting forwards, it severs the vertebral arteries, the eighth 
and ninth pairs, and spinal accessory nerves. A sweep of 
the scalpel across the front of the cord liberates the brain, 
which can be now raised out of the cranial cavity with 

8. The next procedm^e is to place the brain base upwards 
in the skull-cap, the latter being conveniently steadied by 
any simple contrivance. This enables us with less sacrifice 
to the appearances at the vertex, to study the important 
region of the base. In doing so, follow out the instructions 


already given (p. 15), taking the parts in the following 
order : — 

a. Arrangement and appearance of the great vessels at 

the base. 
h. Condition of the membranes and subjacent gyri. 
c. Condition of the various cranial nerves. 
(/. Open up the fissure of Sylvius on either side, tracing 

the large arterial branches upwards ; also examine 

nutrient supply to the basal ganglia at anterior and 

posterior perforated spots. 
e. Strip the base of its membranes, noting adhesions, 

etc. etc. 

9. The cerebrum may now be turned out upon the dis- 
secting tray, with vertex uppermost, and the membranes 
carefully examined and stripped. Take note of the arrange- 
ment of the gyri, the presence of superficial lesions, general 
consistence, and coloiu'. 

10. The clui'a mater and its sinuses at the base of the 
skull may be now examined (p. 8), completing thus the 
examination of the cranial membranes and the external 
aspect of the brain. 



1. Introduce a large section knife into the longitudinal 
fissure, and cut outwards across each hemisphere, about | inch 
above the corpus callosmn, exposing the centrum ovale minus. 
Repeat the same procedure on a level with the corpus 
callosum, exposing the greater centrum ovale, and exhibiting 
the continuation of the transverse strands of this great com- 
missm-al tract with the medulla of the hemisphere. The 
student will now proceed to note the relative and absolute 
amount of grey and white matter, and incj^uire into the con- 
sistence, colour, and other physical qualities of these parts in 
the manner abeady detailed (Chap. IV.). 

72 coause examination of the brain. 

2. By an incision carried from before backwards across the 
fibres of the corpus callosimi, a short distance on each side of 
the median line, the lateral ventricles are opened. To 
expose the structures more fully, divide the corpus callosum 
anteriorly, and whilst gently reflecting it cut backwards, 
with a scissors, through the vertical septum (septum lucidum) , 
which is attached to its under surface, and which, descend- 
ing, divides the lateral ventricles into two cavities. After 
reflecting the corpus callosum, and dividing it across behind, 
note the condition of the following structures seriatim : — 

a. The caudate nuclei or intra-ventricular portions of 

the corpora striata. 

b. Septum lucidum, enclosing betwixt its walls the fifth 


c. Exposed anterior tubercle of the thalamus opticus. 

d. Stria cornea, or exposed part of the taenia semi- 

circularis, coursing between the thalamus and 
corpus striatum. 

€. Fringed margins of the velum interpositum or 

choroid plexus of lateral ventricles. 
/. Fornix, and behind its descending pillars, the fora- 
men of Monro. 

(/. Greneral condition of lining membrane or ependyma. 

3. Divide the fornix anteriorly close to its descending 
pillars, and after examining the subjacent velum inter- 
positum, reflect both structures backwards, exposing the 
thalami optici, separated by the intervening third ventricle, 
and note the condition of the following parts : — 

a. Grey matter forming the boundaries of the central 


b. Anterior, middle, and posterior commissure. 

c. Pineal gland, and its ganglionic extensions, or 


d. Corpora quadrigemina (nates and testes). 

e. Follow out the fornix posteriorly into the descending 

cornua of the ventricles. 


4. On separating the occipital lobes from the cerebellum 
the curved bistoury may, by a little careful dissection, be 
made to cut through both crura cerebri obliquely upwards, 
to meet above the corpora quadrigemina at an obtuse angle 
in the third ventricle in front of the posterior commissure. 

By so doing we separate from the cerebrum the cere- 
bellum, pons, and medulla, together with the corpora quadri- 
gemina and pineal gland intact. This method is certainly 
preferable to dividing the cms at the level of the superior 
cerebellar peduncle, which is so often done at the sacrifice of 
the normal relationships of most important regions. 

5. Separate the cerebral hemispheres by median section 
through the slight remaining connections, and proceed as 
follows : — 

a. Slice one of the hemispheres in a direction parallel 

to the sections already made when exposing the 
centrum ovale. Let numerous sections in this 
direction be made at different planes from above 
downwards through the whole hemisphere, so as 
to expose the structure of the thalamus and corpus 
striatum down to the base. 

b. Slice the remaining hemisphere in a direction at right 

angles to its long axis, or, in other words, from 
above downwards, beginning our sections near the 
olfactory bulb, and proceeding as far back as the 
occipital lobe. 

These sections {a and b) will familiarize the student Avith 
the relationships and coarse structure of the basal ganglia — ■ 
points of considerable moment. Examine, therefore, by aid 
of these sections, the medvdlary tracts forming the internal 
and external capsules, the lenticular and caudate nucleus of the 
corpus striatum, the thalamus opticus and its various regions 
and environment. All these parts may be studied in suc- 
cession, referring to good illustrations, such as appear in the 
last edition of Uuain's "Anatomy." The intimate structm-e 
and relationships of these regions will be dealt with fm-ther on. 


6. Remove the cerebellum from its attachments bj 
dividing the three peduncles, and proceed to examine suc- 
cessively — 

a. Superficial aspect of membranes covering pons and 

h. Condition of blood-vessels and superficial origin of 

the cranial nerves. 

c. Condition of valve of Vieussens, aqueduct, and fourth 


d. By transverse sections expose structure of nates and 

testes, of the pons, its anterior, or motor, and pos- 
terior or tegmental tract, of the medulla oblongata, 
noting the relative dimensions and appearance of 
the olivary bodies, the pyramidal and restiform 

7. Lastly, after examining the lobules and membranes of 
the cerebellum, we divide it into an upper and lower half by 
an incision carried through both hemispheres from behind 
forwards, exposing the central medulla. 

Vertical section — i.e., at right angles to the lamina of the 
cerebellum — will display its foliated aiTangement, and betwixt 
vermiform process and the middle of each hemisphere we 
pass through the plicated corpus dentatum. 



Since the prosecution of researches into the minute anatomy 
of the brain will necessarily lead us into structui'al details of 
infinite diversity (for the complete histology of the human 
cortex cerebri is infinitely more complex than any of our 
classical works on this subject would lead us to conceive), we 
shall have to bestow a predominating share of our attention 
upon cerebral localization. Hence at the outset it is essential 
that we obtain clear and defiuite ideas of the various sub- 
divisions of the brain, both as regards convolutionary 
arrangement and central medullated fasciculi. A knowledge 
of the convolutions and sulci must be immediately acquired, 
and from this superficial topography we learn readily to 
appreciate the relative positions of those smaller areas which, 
from experimental data, have been proved to possess diversi- 
fied functional endowments, as well as those cortical tracts 
which histological inquiry has invested with structural 

It is not within the scope of this manual to give even 
an outline of the internal medullated structure of the brain, 
but it may prove of service here to indicate Professor 
Turner's nomenclature of the convolutions which, from its 
great simplicity and clearness, has found such general favour.^ 
For minute details as to the course, relationships, and devia- 
tions of these gyri, the student is referred to that very 

1 "The Convolutions of the Human Cerebrum Topographically Con- 
sidered," by William Turner, M.B. Lond., F.R.S.E. Ediu. Med. Jnuyn. 
June, 1866, 


excellent translation of Ecker, " On the Convolutions of the 
Human Brain," by J. C. Gralton. From this work, with 
the editor's kind permission, I have tabulated the various 
English and foreign synonyms, as likely to prove of value in 
the fui'ther prosecution of our studies in this department. 
The photographs from Bischoff's work on the Convolutions 
are intended for immediate reference in the laboratory. 


I. Frontal Lobe. II. Paeietai Lobe. III. Occipital Lobe. 
IV. Tempoeo-sphenoidal Lobe. 






Superior Frontal Gyrus 
Middle Frontal G-yru3 , . 

Inferior Frontal Gyrus 

) Separated by the Supero- frontal 
j Sulcus. 

Separated by the Infero -frontal 

Separated by 

the Fissure of 

Separated by the Intra-parietal 

External Orbital Gyi'us. 

Ascending Frontal Gyrus 

Ascending Parietal Gyrus 

Postero-parietal Lobule 

Supra-marginal Lobule 

Angular Gyrus . . 

Third and Fourth Annectant Gyri. 

First Annectant Gyrus. 

Quadiilateral Lobule .. \„ ,,, ,, t, it. 

n a • r» • -i 1 r Separated by the Internal Pa 

Cuneus, or Superior Occipital \ ^, ^ •_;i„i a ,i„„„ 



*j I Fissure. 

Lobule . . 
Lingual Gyrus 

rieto- occipital Sulcus. 

by the Calcarine 

*18. Fusiform Gyrus 




Superior Temporo- sphenoidal Gyrus 
Middle Temporo -sphenoidal Gja-us 
Inferior Temporo-sphenoidal Gyrus ) 
Uncinate Gyrus . . . . j 

Median Aspect of Superior Frontal \ 

Gyrus . . . . . . . . f 

Convolution of the Corpus Cal- ( 

losum . . . , . . , . / 

















by the Calloso-mar- 

ginal Fissure. 

N.B.— At the Base No. 2, 3, 4, indicate respectively the Internal, 
Posterior, and External Orbital Gyri. I need scarcely add that the darker 
and lighter shading of the brain, as shown in the photographs, is well 
calculated to bring into strong relief the moi'e distinctive areas and divisions 
of the surface. The relative areas occupied by difierent lobes over each 
aspect of the hemisphere are thus better displayed than by outline figures, 
whilst the distinctive character of the gyri is not interfered with. 

» Neither of these terms is included in Turner's nomenclature. They are adopted 
by Ecker. 



SuPERioB Feo^jtal Gyeus. 

Erste oder obere Stirnwindung . . . . {Ecker.) 

Etas'e frontal superieur ou troisiSme . . ) ,^ ^- , ^ \ 

° \ [ {Gratiolet.) 

Pli de la zone externe . . . . . . . . ) 

Supero-fi'ontal gyrus . . . . . . . . {Rnxlei/.) 

Middle Feontal Gyeus. 

Zweite oder mittlere Stirnwindung . , . , {Eclrr.) 

Etage frontal nioy en .. .. .. .. {Gratiolet.) 

Medio-frontal gyras . . . . . . . . [Huxlcij.) 

Infeeior Feontal Gyeus. 

Dritte oder untere Stirnwindung .. .. {Ecker.) 

Pli frontal infferieur ou premier ou etage 

surcilier .. .. .. .. ,. {Gratiolet.) 

Infero-frontal gyrus . . , . , . . . {Huxley.) 

Ascending Feontal Gyrus. 

Vordere Centralwindung .. ., .. {Rusckke.) 

Processi enteroidei verticali di mezzo (anterior 

part) . . . , . . . . . . . . {Rolando.) 

Circonvolution transverse parietale anterieur (Foville.) 

Premier pli ascendant . . . . . . . . {Gratiolet.) 

Antero- parietal gyrus . . . . . . , . {Huxley.) 


Erste obere Schlilf enwindung . , . . ( Wayiier. ) 

Antero-temporal . . . . . . . . {Ha.dcy.) 

Pli temf)oral superieur ou.. .. ..\ 

Pli marginal posterieur ) {Gratiolet.) 

Gyrus temporalis superior sive infra marginalis {Huschke.) 

Middle Temporo- sphenoidal Gyrus, 

Zweite oder mittlere Schlafenwindung 
Mittlere Schlafenwindung 

( Wagner.) 



Pli temporal moyen ou partie descendante du 

pli courbe . . . . . . . . . . {Gratiolet.) 

Medio-temporal gyrus .. .. ... {Huxhij.) 

Inteeiok Tempoho- sphenoidal Gyeus. 

Dritte oder imtere Schlaf enlappenwindung i 
Gyrus temporalis tertius sive inferior . . ' 

Lingual Lobule. 

TJntere innere Hinterhauptwindungsgruppe 
Lobulus LinguaUs — Zungenlappchen 
Gyi-us occipito -temporalis mediaUs . . 

Fusifoem: Lobule. 

Untere aussere Hiuterhauptwindungszug . . 
Gyrus occii^ito-temporalis lateralis . . 
Spindellappchen . . 

Lobulus fusiformis — Spindelformiges 

Ascending Pakietal Gyrus. 

Ilintere Centralwindung 
Processi enteroidei verticale di mezzo 
(posterior segment) . . . . . . ■ 

Cu'convolution tranverse medio-parietale . . 
Deuxieme pli ascendant 
Postoro -parietal gyrus 

PosTERO- Parietal Lobule. 

Oberes Scheitellappchen 

Gyrus parietalis superior 

Lobule du deuxieme pli ascendant . . 

Erste Sclieitellappen-windung 

Obere Sclieitelbeinlappen 

Obere innere Scheitelgruppe . . 

Supra- Marginal Lobule. 

Dritte Scheitellappenwindung 

Unterzug aus der hintern Centralwindung ) 

ScheitelhiJckerlappclien . . . . . . ' 

PU marginal superieur 

Erste oder vordere Sclieitelbogenwindimg . . 

Angular Gyrus {Huxleij). 

Unteres Scheitellappchen (posterior part) . . 
Zweite oder mittlere ScheiteUappenwindung 
Auf steigende "Windung zum hintern aussem j 

Scheitellappchen und 
Hinteres iiusseres Scheitellappchen 


( Wagner. ) 

{Panschand EcJcer.) 


{Fansch and Ecker.) 





{ Wagner.) 
[Huschke .) 



{ Wagner.) 




Zweite odei' mittlere Scheitelbogenwindung [BiscJioff.) 

Pli courbe .. .. .. .. ,. {Gratiolet.) 

Quadrate Lobule (Ricrley.) 

Lobule quadrilatere 




Convolution of Corpus Callosum. 

Gyrus Fornicatus — Bogenwulst 
Gyrus cingiili Zwiuge or Cingula 
Callosal gyrus . , 
Circonvolution de I'ourlet 
Processo enteroidio cristato . . 






Uncinate Gyrus {Huxley — Turner). 

Gyrus Hippocampi 
Subiculum Cornu Ammonis 
Circonvolution a crochet 
Pli Unciforme . . 
Temporal moyen interne 
Lobule de riiippocampe 

Dentate Gyrus. 

Coi-ps godronne . , 

CuNEUS {Burdach, Turner, etc.) 

Zwickel . . 

Erste oder Hinterhauptlappenwindung 

Oberer Zwischensclieitelbeinlappen . . 

Lobule occipital 

Internal occipital lobule 

First Annectant Gyrus. 

Erste oder obere HinterhauptAvindung 

Oberer Zug der liintern Ceutralwindung . . 

Erste obere Hinteiiappenwindung . . 

Obere innere Scheitelbogenwindung 

Fir^t external annectant gyrus 

Pli de passage superieur externe . . . . ) 

Pli occipital superieur . . . . , . . . j 

{ Vicq d'Azi/r.) 


{Gratiolet, etc.) 

{Gratiolet. ) 

{ Wagner.) 


Second Annectant Gyrus. 

Zweite oder mittlere Hinterliauptwindung. . 
Zweite mittlere Ilinterlappenwindung 
Pli occipital moyen . . . . . , , , ) 

Deuxieme pli de passage externe . . . . j 

Gyrus occipitalis medius 

( IVac/ncr.) 




Medio -occipital and second external an- 

nectant .. .. .. .. .. {Huxley.) 

Third and Fourth Annectant Gyei. 

Dritte oder untere Hinterhauptwindimg . . {Echer.) 

Dritte untere Hinterlappenwindung .. {Wagner.) 

Pli occipital inferieur . . . . . .\ 

Troisieme et Quatrieme pli de passage ^ {Gratiolet.) 
exteme . . . . . • . . . . ' 

Gyrus occipitalis inferior .. .. .. (Pause h.) 

Lobes of the Ceeebeum. 


Parietal . . 
Temporo-splienoidal . . 

Occipital . . 
Central Lobe 

= Scheitellappen. 

= Stiralappen. 

= ScMafenlappen.i 

Keilbeinlappen . ^ 

Schlafen-KeQbeinlappen. ^ 
= Hinterhauptlappen. 
= Zwiscbenlappen. 





1 •= Temporal 


■* - Temporo-sphenoidal. 

Fig. 4.— convolutions of the cerebrum, as seen 

AT THE VERTEX (after Bischnf). 

AT THE MEDIAN ASPECT (after Bischof). 


^ \ 







AT THE BASE (nficf BUchof). 


ASPECT (after Bischof). 


G 2 



Prior to considering in detail the methods adopted for the 
minute examination of the brain, the student should be fully 
impressed with the conditions which render many of the 
processes applicable for other tissues quite inadmissible here. 
The consistence of cerebral tissue is such that cutting fine 
sections by the hand, either with an ordinary razor or the 
Valentin blades, cannot be adopted with any hope of success, 
since the finest sections thus obtainable are useless for micro- 
scopic examination, which requires sections of extreme 
tenuity. These extremely fine sections can only be obtained 
by aid of the screw microtome from portions of hardened or 
frozen brain. In order to modify the consistence of the 
tissue so as to adapt it for this purpose, various methods of 
hardening by chromic acid, chromic salts, alcohol, picric acid, 
and osmic acid have been devised. The results which have 
accrued from these methods have been most valuable, yet they 
are uniformly open to the serious objection that this altered 
consistence induced by the is obtained at the expense 
of modified structure and altered relationships. Amongst the 
most serious drawbacks to the use of corrugating reagents in 
the examination of the brain are the following : — 

A considerable expenditure of time (from three to eight 
weeks being requu-ed for satisfactory hardening). 

The process itself is extremely tedious, and often unsatis- 
factory in its results, as evidenced by experienced 


It requires a considerable amount of practised manipula- 
tion, and exposes the tyro to numerous disappoint- 
ments and failures. 

The shrinldng of tissue is a most objectionable feature in 
the cortex, often a,mounting to one-half the original 

The normal wealth of structm-e is greatly modified; in 
this respect it is infinitely surpassed by fresh methods. 

The absolute and relative depth of the various layers of the 
cortex is subjected to most objectionable modifica- 

It affects the cortex of different animals to a different 
extent, thus interfering with comparative investiga- 
tions. Many pathological and normal aj)pearances 
are wholly obliterated by the " hardening processes." 

Of all tissues, that of the brain, from its extreme delicacy 
and susceptibility to rapid post-mortem change, is the tissue 
which, above all others, demands a rapid, ready, and fresh 
method of preparation, the employment of indifferent media, 
and the restriction of all corrugating reagents. No method 
as yet adopted can surpass for elegance, expedition, and 
certainty the freezing method ; and, above all, it is f/w 
method which should be chiefly trusted to in all exact 
anatomical and pathological research. Yet, whilst we claim 
for the fresh freezing methods such self-evident advantages 
over the older process, the long series of objections to the 
latter tabulated above must not induce us to blind our eyes to 
the fact that it also possesses its own advantages and cannot 
be dispensed with. Most of our classical descriptions of 
cerebral structure refer to brain which has been subjected to 
these methods of preparation ; and few indeed, far too few, 
are the delineations of the minute structiu-es of the cortex 
and medulla in the perfectly fresh brain. Hence, as a 
question of simple comparison between the results of different 
observers and our own, the hardening methods must still be 
adopted. There are numerous other very apparent reasons 


■why we cannot afford to dispense with the older processes, 
and the student is therefore recommended, whilst guarding 
himself from its fallacies, checking or confirming its results 
by the fresh methods, to make himself equally familiar 
with the practical details of both methods, and to learn to 
discriminate the special work to which each process is more 
particidarly suited. 

Two Distinct Methods of Freezing. — Just as we find 
amongst the list of reagents used for hardening brain, some, 
and especially osmic acid, far less open to the objectionable 
qualities possessed by alcohol and chromic acid, inducing in 
fact very slight alteration in the relationships and bulk of 
the tissue, so do we find the different methods of freezing- 
possess their o^vn distinct and relative merit. This fact, 
which it is all-important to recognize when dealing with 
structures like the brain, has been wholly misunderstood or 
overlooked by all authorities who have written upon the 
subject, and we constantly find histologists grouping the 
freezing by ice and salt with the ether freezing method, as 
though both were equally suitable for all tissues alike. 
Now the fact is that the ordinary ice and salt freezing micro- 
tome is entirely useless for brain-structures, except after a 
modification of the latter by a process of hardening, and 
hence no longer a fresh method. Upon the other hand, the 
ether freezing microtome is applicable to all tissues having a 
consistence not above that of the liver, but is pre-eminently 
adapted for nervous tissue. I find it necessary to insist upon 
this point, since it seems generally understood that the ice 
and salt mixture is suitable for freezing brain. Prior to the 
introduction of freezing by ether, practised liistologists con- 
stantly complained of the impossibility of obtaining fine 
sections of brain by freezing, the objections being that hard 
spiculse formed within its structm^e, and tore it up on cutting 
through it. Yery lately this fact has been again asserted by 
a skilled manipidator,^ who has even introduced a method of 

^ Vide Hamilton in the Journ. Anatomy and Fhys. vol. xii. p. 259. 


combined hardening and freezing to overcome this very 
unsatisfactory action. Hitherto, therefore, it may be 
asserted that the use of the ice and salt microtome has 
failed for the purposes of the cerebral histologist ; and 
although by further development it may be rendered sub- 
servient to his purposes, for the present the freezing of 
brain-substance must be uniformly pursued upon the ether 
freezing microtome. The methods available for the exami- 
nation of the minute structure of the brain are as follows: — 

Hardening processes for sections to be cut on the imbedding 

Hardening processes for sections to be cut on the freezing 

Fresh process by means of the ether freezing microtome. 
Fresh process by a modified teazing and staining. 
Fresh process by ordinary teazing with dissociating 


We will preface our account of these methods by a brief 
outline of the more useful microtomes used. 

microtomes for imbedding and freezing. 

Microtomes for Imbedding. 

Amongst the various instruments devised for cutting sections 
of hardened tissues are the microtomes of Henson, Rivet- 
Leiser, Brandt, Roy, His, Ranvier, Stirling, and Rutherford. 
The instrument more generally used in this country is that 
devised hy Stirling, and its modification by Rutherford, 
either of which instruments will prove satisfactory to the 
student ; whilst that of Ranvier may be employed where it 
is desirable to grasp the microtome in the hand, or immerse 
it during section-cutting in water or spirit. 

The principle adopted in Stirling's instrument has been 
very generally followed. In the last three named, the body 
of the instrmnent consists of a metal tube or hollow cylinder, 
fixed to a table by some simple arrangement, or held in the 
hand in an upright or vertical position. A fine micrometer- 
screw works into the lower end, driving through the hollow 
of the cylinder a closely-fitted piston-plug, which in its turn 
propels the mass in which the tissvie to be cut is imbedded. 
The essential portion of this form of microtome, therefore, 
is the body or cylinder, and the powerful micrometer-screw. 
The body, or cylinder, includes the following divisions: — 
the hollow, or " well " of the cylinder ; its closely-fitted 
plug, or "piston ;" the upper smooth and levelled extremity, 
or " section-plate ;" the lower extremity forming a " female 
screw " for the reception of the micrometer-screw. It is far 
preferable that the cylinder should be clamped by projecting 


arms and screw to a firm support as a table. In choosing a 
microtome for cutting sections of an imbedded tissue tbe 
student must be guided by the following considerations. 
The instrument should be strong but compact, and not 
weighty or cumbersome ; it should possess appliances for 
fixing it firmly and immovably to a supporting ledge or 
table. Its well should be at least one inch in diameter, and 
the oval section is to be preferred to the circular well, since 
this form of well wholly prevents rotation of the imbedded 
mass. The section-plate should be absolutely level, perfectly 
smooth, polished, and show no irregularities of sm^face or 
indentations around the margin of the well, which otherwise 
would ruin the edge of the blade. The micrometer-screw 
should be of powerful leverage, work evenly, easily, and 
without the slightest " loss of time," and have a pitch of at 
least fifty threads to the inch. Allowing his judgment to be 
guided by the above rules, the student cannot err greatly in 
his selection of a microtome for ordinary work and the smaller 
class of sections. A few words on the different varieties of 
instruments used may prove of service to the student. 

1. Stirling's Microtome is a compact, strong, and 
admirably finished instrument, embracing all the qualities 
essential for the section-cutting of hardened preparations. 
For small sections no better adapted form has yet been 
devised, and for his earliest attempts in cutting hardened 
brain the student is recommended to secui-e this form of 
microtome. This instrument is made to be clamped to a 
table, so that both hands are free for section-cutting. 

2. Ranvier's Microtome, — This is a smaller but very 
useful instrument. It has, however, to be held in the hand, 
a defect which is in part counterbalanced by the ease with 
which it can be immersed in spirit, and sections cut whilst so 
situated. It is well adapted for sections of spinal cord and 
the large nerve-trunks. 

3. Rutherford's Microtome. — This, which is a modified 
form of Stirling's microtome, is a most valuable instrument, 



as it is well adapted for section-cutting of imbedded or of 
frozen tissues. It will be described fully in the section on 
freezing microtomes. It has the disadvantage of being some- 
what cmnbersome as compared with the smaller " Stirling," 
whilst at the same time as a freezing instrument for nervous 
tissues it is excelled by the ether freezing microtomes. The 
latter remark does not apply to other animal tissues. 

4. Roy's Microtome^ consists of an inner vertical brass 
plate covered with a layer of cork, and sHding by vertical 
movement within an outer brass frame-work. A glass rod 
of horse-shoe form is fixed horizontally by its extremities 
into the brass frame, upon which the blade glides during 
section-cutting. The imbedding mixtm^e is first cast in a zinc 
mould, the tissue being placed in it, and when hardened, the 
mass is fixed upon the cork plate. The vertical movement of 
the brass slide is obtained by propulsion from a fine-threaded 
screw fixed below. A simple tubulature, adapted to the 
instrument, allows spirit or water to be blown upon the mass 
when requii-ed. It has been stated that tissues may be cut 
fresh with this microtome after freezing by the direct appli- 
cation of the ether spray. The student must be cautioned 
against any such attempt; for all tissues alike, the dii"ect 
application of ether is to be deprecated, and for brain and 
spinal cord and all nervous structures such a method is 
wholly inadmissible. Eoy's microtome has the disadvantage 
of not being readily fixed to a table during manipulation. " 

5, Schiefferdecker's Microtome is described here as 
appUcable to hardened preparations, although it is essentially 
constituted to dispense with the method of imbedding. The 
object to be cut is fixed within a hollow brass cylinder by 
means of a clamp upon its surface worked b.y two screws. 
In lieu of a propelling screw below, as in other microtomes, 
a circular plate forming the section-plate can be elevated or 
lowered by the revolution of an outer concentric plate which 

1 Described and figured in the Journal of Physiology, vol. ii. No. 1. 

2 Tlie instrument is made hy Mr. Gardner, South Bridge, Edinburgh. 


turns upon a screw. The student will find this microtome 
figured and described in the Qiiarterhj Jownial of Micro- 
scopical Science for January, 1877. 

6. Microtome for Slicing through Whole Hemi- 
sphere of Human Brain. — The instrument which I have 
had made for this purpose at the West Riding Asylum consists 
of a heavy brass cylinder, 3 inches deep and 8 inches in 
diameter, closed in below, where, however, its cavity com- 
municates centrally with that of a small secondary cylinder — 
the screw-socket. In this socket, which is 4 inches deep, 
works a powerful and finely-threaded screw, having a milled 
head 3 inches in diameter. A flat, circular brass plate, 
accm'ately fitted to the interior of the upper cylinder or well, 
is raised or lowered by means of the screw, the movement 
being communicated to the imbedding mass, which rests 
upon it above. The section-plate is constituted by the 
projecting rim of the cylinder above, and is mathematically 
level and smoothly polished. The microtome rests upon the 
iron collar of a powerf id tripod stand. A zinc tray, 2\ inches 
deep and 26 by 17 inches, can be adapted to the microtome- 
cylinder, so that sections may be cut under water with the 
greatest ease. 

Microtomes for Freezing. 

1. The Ether Freezing Microtome (Bevan Lewis). 
— This microtome was described in the Journal of Anatomy 
and Physiology for April, 1877, but has been modified and 
improved in several resjDects by the present maker. ^ It 
consists essentially of «, the body ; h, the freezing chamber ; 
c, the section-plate. 

a. The Body. — This is really a modified " Stirling micro- 
tome," and forms the lower half of the instrument. Through 
its central aperture works a brass plug, driven by a strong 
but fine micrometer-screw. In my own instrument, the pitch 

' The improved microtome may be obtained of Mr. Gardner, 45, South 
Bridge, Edinbvu'gh, who, at a minimum of charge, has fully perfected the 
woi'kmanship. Descriptions of the microtome may be also found in Brain, 
October, 1878; and in Dr. Stirling's " Text -book of Practical Histology." 



of the screw is fifty tlireads to the inch ; the screw has a diameter 
of ---inch, with a milled head, Ih inches across. A coarse 
screw secures the body of the instrument to a table. 

b. The Freezing Chamber. — This consists of a zinc cylinder, 
closed above and below, pierced on either side by a large 

Fig. 8.— The Ethee Freezing Microtome. (Original Model.) 

aperture, to allow of the admission of the nozzle of the spray- 
producer, as well as to permit free evaporation of ether, 
much of which, however, condenses on the bottom, and is 
conveyed off by the bent tube into a bottle attached to it. I 
have found it convenient to have the zinc cylinder quite 
21 inches wide by Ih inches deep, and the aperture in the side 
of the instrument f -inch in diameter. 

The cap or plate closing the cylinder above, I term the 
freezing plate, and upon it rests the tissue, whilst the ether 
spray plays upon the plate from below. The freezing- 
chamber is thus rapidly reduced in temperature, and to pre- 
vent conduction by means of the section-plate, it is well to 
have a free interval all around the freezing cylinder, securing 
it from actual contact with the section-plate. 



c. The Section-Plate. — This is made either of metal or of 
plate-glass, drilled with a central aperture, through which the 
freezing chamber glides. If of metal, it should, by prefer- 
ence, be made of a smooth, polished zinc plate, about -f -inch 
thick, supported by strong vertical arms upon the body of 

Fia. 9. — TnE Ether Freezing Microtome. (New Form.) 

tlie instrument. I would strongly advocate the use of the 
plate-glass, since it renders the movements of the knife abso- 
lutely free and easy ; whilst, on the other hand, the blade 
never gets injured from scratches, which, sooner or later, 
invariably appear upon a metal section-plate, and wliich turn 
or indent the edge. 


To recapitulate — the various divisions of our microtome are 
as follows : — 

a. The "body," comprising the "well," the "piston," 
and the " micrometer-screw." 

h. The " freezing chamber," fitted with sloping false- 
bottom, an " exit-tube," and capped by the " freezing 

c. The "section-plate," drilled for the passage of the 
" freezing chamber." 

The Freezing Medium. — The best anaesthetic ether 
should be employed. I find it more economical than 
methylated ether. The ordinary Richardson ether spray is 
the more generally used instrument ; but a further improve- 
ment has been introduced by the maker, in the form of a 
lateral support for the ether-spray apparatus, communicating 
with a bellows worked by the foot. In this form, the spray 
adjustment is easy, and the hands are both free for section- 
cutting and manipulation, and thus a most substantial and 
valuable improvement has been obtained, with but a very 
trifling addition to the cost of the microtome. 

2. Rutherford's Freezing Microtome.^ — This instru- 
ment consists of a brass section-plate, with a central aperture 
leading into the interior of a vertical tube, in which a plug 
is fitted to move upwards or downwards by means of a fine 
screw. The cylinder is surrounded on all sides by a metal 
box covered with gutta-percha, which holds the freezing 
mixture of ice and salt. An exit tube allows of the escape 
of water. The whole instrument can be securely clamped to 
a table. The method of using this instrument for freezing 
is as follows : — The plug is first unscrewed and oiled, so as 
to prevent its fixture clming the process of freezing. The 
tissue to be frozen and cut is first immersed in a thick 

' "A New Freezing Microtome:" Monthly Micros. Journnl, vol. x. 
p. 185. "Outlines of Practical Histology : " by William Rnlherford, M.D. 
2ncl edit. p. 164. 



solution of gum for some hours, " in order that the gum may 
permeate every part of the tissue, and prevent the formation 
of a crystalline condition within the frozen tissue." If the 
tissue has been previously immersed in alcohol, all traces of 
the latter shoidd be first removed by soaking in water. 
Equal parts oi finely-powdered ice and salt are now placed in 
the freezing box and stirred around the well, whilst the latter 
is filled with the solution of gum. As the gum freezes 
around the periphery, the tissue is plunged into it, and held 

Fig. 10. — Exttherford's Freezing Microtome. 

until it is fixed by the advancing ice. Care should be taken 
to keep the exit-tube of the box open, so as to allow of free 
escape of water, and to close the freezing box by a weighted 
strip of cork, to prevent the entrance of heat and to exclude 
the salt of the freezing mixture. By this method the freez- 
ing process may be accomplished in from ten to twenty 
minutes. All delicate tissues require a sipecial prejmrofion to 
adapt them for this process, otherwise the water which they 
contain freezes into hard icy spicules, whicli tear the structure 


when it is cut. Dr. Pritchard has therefore recommended a 
prior immersion of the tissue in thick solution of gmn-arabie, 
using the same solution as the imbedding material. 

Quite recently, Dr. Hamilton whites as follows with regard 
to freezing the nervous structures, and especially brain, by 
means of the ice and salt microtome. " It was found that 
the crystals of ice so broke up the delicate nervous tissue as 

to render it totally useless for minute examination I 

attempted two years ago to modify the method of freezing, 
but without success, and accordingly gave it up as an almost 
hopeless task." ^ 

According to this statement, the writer's exi^erience tallies 
wholly ^^'ith my own ; and f idly acquainted as he was with the 
methods of steeping tissues in gum prior to freezing which 
were adopted by Dr. Eutherford, it is clear that this method 
of freezing was not applicable for fresh brain, although it 
answers admirably in the case of firmer and less delicate 
textm-es. Dr. Hamilton overcame the difficulty of freezing 
brain with the ice and salt mixture, but at the expense of 
sacrificing the fresh for the chrome hardening process, a 
special procedure being requisite, which will be considered 
further on. Now it is after such considerations that we 
recognize the undoubted superiority of the ether freezing 
microtome for nervous structures. It has been stated that the 
ether freezing process has been entirely superseded by the 
introduction of ice and salt, or other mixtures, which by con- 
stant refrigeration keep the mass in a frozen state for houi's. 
All practical manipulators, however, know that this is, as 
regards nervous structures, a fallacy and a blunder, since the 
very excellence of the ether freezing method depends upon 
the fact that it can be checked at any stage and renewed 
when required. This is the all-important consideration in 
freezing brain, for beyond certain limits it is frozen into a 
hard icy solid, which at once blunts or tm-ns the edge of the 
blade ; but with the ether-spray this stage need never be 
attained, and a consistence is obtained admirably adapted for 

' Jonrti. of Aiuif. and riiijs. vol. xii. p. 257. 



section-cuttiug. The student should therefore bear in mind 
that the constant cqyjjiication of a freezing mixture to fresh brain 
cannot but result in failure for section-cutting ; and that bi/ use 
of the ether process the prior jnrparation of brain by hardening, 
or by immersion and saturation in mucilaginous fluids, may be 
wholly dispensed irith. 

3. Williams' Freezing Microtome.^ — In this instru- 
ment a wooden tub containing a freezing mixture of ice and 

Fig. 11.— Williams' Freezing Miceotome. 

salt is covered by a glass lid, the frame of which is seciu-ed 
by a clamp screw. An upright brass conducting-bar passes 
from the interior of this box through a central apertm-e in 
the lid, and to the extremity of this bar is screwed the 
circular brass plate which supports the tissue to be frozen. 
The arrangement of the knife is peculiar, since it is fixed in 

1 This instrument is made by Messrs. Swift & Son, University Street, 


a triangular frame, and can be either raised or lowered by 
the screws which are adapted to the frame. There are three 
additional plates for supporting the frozen object, and a brass 
cup for holding substances, which are fixed in cacao-butter 
or paraffin. 

The box is filled with equal parts of pulverized ice and 
salt, care being taken to prevent the mixture touching and 
so fixing the cover. After the cover is replaced and screwed 
down, the substance to be cut is placed on the central brass 
plate, surrounded by a little solution of gum, and the appa- 
ratus is covered with baize to facilitate freezing. When 
frozen, raise the blade, and after the fii'st cut across, proceed 
as follows : — Each end of the razor must be presented to the 
surface of the section, and exactly levelled by means of each of 
the hack screws. If the large back screw be now tiu-ned, the 
blade can be lowered to any required extent ; and since a 
complete revolution of the screw gives us a section -p^-jy-inch 
in thickness, and the screw-head is graduated into sixths — a 
movement through sixty degrees gives us a section ^-^p-inch 

The special featm'es of this microtome, in which it differs 
materially from others already described, consist in an 
arrangement whereby the blade and not the imbedded or 
frozen tissue becomes the movable part of the instrument ; 
and the edge of the knife is only brought into contact with 
the substance to be cut. 




In subjecting the brain to the agency of hardening reagents 
certain important conditions should always be kept in mind, 
as they are essential to success. The conditions are as 
follows : — 

a. The reagent should act equably upon all portions of 

the tissue. 

b. The requisite consistence must be acquired at the 

expense of the minimum of alteration and shrinking 
of tissue. 

Now the fii'st of these conditions can only be obtained by 
ensuring a thorough saturation of the tissue throughout, so 
that the fluid permeates rapidly to the central or deepest 
portions of the mass. It is evident that the surface of the 
tissue being bathed in the reagent will be more actively 
affected by the latter than the more distant parts within, 
and thus arises the danger of a too rapid hardening of the 
exterior, which forms a mechanical impediment to the per- 
meation of the deeper structures by the surrounding fluid. 
Always, therefore, take the precaution to ensure a free and 
rapid permeation of the tissues by the fluid. 

Again, if any portion of the surface is in close contact with 
the side of the containing vessel, it necessarily is less affected 
than the surface bathed in the fluid. The mass should there- 
fore be so suspended that it is equably bathed all around in the 


Another important consideration is that of temperature, for 
under the most favourable cii'cumstances the central or deeper 
structures must remain far less subject to the action of the 
reagent than the exterior of the mass, and are, therefore, the 
parts which most readily succumb to putrefactive changes. 
The larger the mass, therefore, to be hardened, the moi'e 
difficult is it to prevent central decay, whilst an elevated 
temperatui'e induces the same result. The tissue to be 
hardened should therefore be of moderate hulk as compared 
with the fluid in which it is immersed, and the preparation 
should be kept in a cool spot, or better still, in an ice safe. 

The next condition we have named, and which it is equally 
important to secure, is that a minimum of shrinking of tissue 
should result from the action of the reagent, and it is a well- 
known fact that all the reagents used for this purpose will, if 
employed in too concentrated a form, ruin the preparation 
by inducing extreme shrinking and brittleness of the mass ; 
whilst again, some of these reagents are far more reliable than 
others, and less open to these disadvantages. The more 
commonly used hardening reagents are — osmic acid, Miiller's 
fluid, solutions of the chrome salts, chromic acid, picric acid, 
methylated spuits, and alcohol. Now this list represents their 
relative value as hardening reagents for nervous tissues, osmic 
acid and Miiller's fluid standing at the head of the series as 
the most valuable and least injurious in their action, chromic 
acid and alcohol occupying a far less prominent position in 
the scale. In order therefore to secm^e the tissue from 
injurious shrinking, employ in preference to the others the 
reagents noted at the head of the list, and use the weakest solu- 
tions compatible with safety to the tissue, commencing with the 
weakest, and gradually augmenting the strength of the solu- 
tion, or, later on, even replacing by the more astringent 
reagents. Let us now detail throughout the process recom- 
mended to the student for his first essay in chrome-harden- 
ing, subsequent to which it will be useful to briefly dwell 
upon the various modifications of the process adopted by 



1. Miiller's Fluid and Potassium Bichromate. — 
Excise a portion of the ascending frontal or parietal convolu- 
tion of human brain, cutting across its length so as to remove 
about an inch of the convolution along with its central and 
deeper medulla. Lightly cover it all round with a little 
cotton-wool, and immerse it in from two to three ounces of 
methylated spirits contained in a four-ounce stoppered bottle. 

Label the bottle with the name of the specimen and date, 
placing it in a cool cellar or ice safe. 

In twenty-four hours pour away the spirit, replacing it by 
four ounces of Miiller's fluid, the preparation being sur- 
rounded as before by cotton-wool to ensure it being bathed 
upon all sides alike by the fluid. 

Let the bottle stand in a cool spot, and in three days 
replace the fluid by a fresh quantity. At the end of one 
week the fluid should be again renewed, or, preferably, a. 
weak solution of potassium bichromate substituted (2 per 
cent.). At the end of the second week a solution of the latter 
of double the strength may be added ; and if at the termina- 
tion of the third week the mass is still pliable, and of the 
consistence of ordinary rubber, it is as yet unfit for section- 
cutting, and the reagent should be rej)laced by a solution of 
chromic acid. 

In these later stages the chromic acid expedites the process 
without producing the extreme shrinking of tissue which 
ensues if it be used at earlier stages. For the same reason 
even absolute alcohol may at this stage be employed, but 
although valuable in the preparation of the spinal cord, it 
cannot be equally well recommended for the cortex of the 
hemispheres. By the above process our preparation will have 
attained the requisite consistence within a period of from four 
to eight weeks. It will be observed that in the various steps 
of this process the conditions previously emphasized as essential 
to success are obtained. Thus the early immersion in methy- 


lated spirit abstracts all superfluous fluid from the brain aud 
its vessels, and entering its substance by its afiinity for water, 
aids in tbe rapid permeation of the mass by the chromic solu- 
tion in the next stage. The more prolonged action of the 
spirit would, however, prove highly detrimental. Again, the 
cotton- wool ensures an equable distribution of fluid around, 
whilst the comparative bulk of the mass and fluid in which it 
is immersed, and the temperature to which it is exposed, 
provide for equable hardening to its deepest structures, and 
ensure it against decomposition. The gradual increase in the 
potency of our reagents from Miiller's fluid to the strong 
solutions of chrome is also an adjunct to the hardening of 
the central portions, whilst we also expedite the process 
of hardening. 

2. Potassium Bichromate and Chromic Acid 

(Eutherf ord) . — Place small portions of the cerebrum in 
methylated spirit for twenty-four hours, observing the same 
precautions as to relative bulk of the preparation and the 
reagent, covering with cotton-wool, and leaving in a cool 
place. Replace the spirit by a mixture of potassium bichro- 
mate and chromic acid. The proportions may advantageously 
be varied, according to the condition of the structure to be 
hardened ; but the solution recommended by Rutherford, and 
which answers well, contains 1 gramme of chromic acid and 
2 graromes of potassium bichromate to 1,200 c.c. of water. 
Change at the end of eighteen hours, and then once a week. 
Should the tissue not be sufficiently tough for cutting at the 
end of six weeks, place it in a ^ per cent, solution of chromic 
acid for a fortnight, and then in rectified spirit. 

3. Iodized Spirit and Potassium Bichromate (Betz). 
— Large portions of cerebrum may also be placed for a few 
hoiu-s in methylated spirit, tinted of a light sherry-brown by 
tincture of iodine. Add fresh iodine solution as the colour 
fades. In one or two days remove the pia mater, and return 
the preparation to the solution, adding to the latter haK its 
bulk of fresh iodized spirit. After the lapse of another period 


of two days, replace tlie solution by iodized alcohol (alcohol 
70° — 80°) tinted of a sherry-brown by a tincture of iodine. 
In from two to three days it should be transferred to a 4 per 
cent, solution of potassium bichromate until sufficiently 
hardened for section-cutting. Shoidd a brown deposit form 
over its surface during the process, let it be well washed, and 
a fresh solution used. When hardened, these preparations 
may be kept permanently in 0*5 per cent, solution of bichro- 
mate. If the cerebellum is to be hardened, the segments are 
at once placed in the iodized alcohol, adding fresh iodine 
frequently as the colour of the solution j)ales. Remove, the 
pia mater on the second or third day, and in a week transfer 
to pm'e methylated spirit for twenty-fom^ hours, and finally 
harden it in a '5 per cent, solution of potassium bichromate. 
This process, recommended by Prof. Betz, is not suitable 
for examination of the cortex, as the iodized spirit is injurious 
to the after-processes of staining. It is, however, especially 
suited for obtaining large sections through the hemisphere, 
the whole cerebrum and cerebellum, when sliced evenly across 
into segments |-inch thick, being most satisfactorily hardened 
throughout. For the minute examination of the cortex 
methods 1 and 2 are pre-eminently to be preferred. 

4. Miiller's Fluid and Ammonium Bichromate 
(Hamilton). — This method is especially applicable to large 
segments of the brain, and is much to be preferred to the 
process by chromic acid, 5. The brain is sliced completely 
through into segments about one inch thick. Each segment 
may now be placed in a large vessel, such as a brain-prepara- 
tion jar, padded with cotton-wool, and containing a com- 
paratively large bulk of the solution, which consists of three 
parts of Midler's fluid to one part of methylated spirit. A 
refrigerator or ice safe should invariably be employed to pre- 
clude decomposition, and the pieces should be turned over 
occasionally in the solution. In about three weeks they 
may be transferred to a solution of ammonium bichromate 
(•25 per cent.). At the end of the fom'th week replace by a 


1 per cent, solution of the same salt, and the following week 
by a 2 per cent, solution, in which they remain until fit for 

5. Chromic Acid Solution (Lockhai-t Clarke). — The 
convolutions of the cerebrum and cerebellum were by tliis 
process hardened in a 0*25 per cent, solution of the crystallized 
chromic acid — a stronger solution rendering them far too 
brittle for section-cutting. As stated above, chromic acid is 
not adapted for hardening so satisfactorily and uniformly as 
the chrome salts. This method, emploj^ed by Lockhart 
Clarke, has been superseded by more reliable methods, as 
are also the processes recommended by Stilling, Kolliker, 
Hanover, and Yan-der-Kolk, for the hardening of nervous 


It has been already stated that osmic acid (OsO^) is one of 
the most reliable agents for hardening the brain and other 
nervous textures which we possess. Prof. Sigm. Exner, of 
Vienna, has therefore devised a method whereby small por- 
tions of the brain may be prepared for section-cutting by 
means of this reagent.^ 

Exner's Process. — A small portion of brain, not ex- 
ceeding one cubic centimetre in size, is placed in ten times 
its volume of a solution of osmic acid (1 per cent.). The 
solution should be replaced by a fresh reagent of the same 
strength after the lapse of two days — a proceeding which may 
be advantageously repeated at the end of the fourth day. In 
from five to ten days the piece is usually stained throughout, 
for this reagent has the valuable property of hardening and 
staining simultaneously. The hardened brain is then washed 
in water, plunged for a second in alcohol to facilitate the 
imbedding, and sections are cut in the iisual way in an 

' "Zur Kenntniss vom feineren Baue der Grosshimrinde." Aus dem 
Ixxxiii. Bande der Sitzb. der k. Akad. dcr Wissensch. iii. Abth. 1881. 


ordinary microtome. The subsequent treatment of these 
sections will he given later on, when methods of staining and 
mounting are considered. It is only necessary here to remind 
the student that osmic acid is really a most valuable agent in 
the investigation of brain-structm^e, and he shordd avail him- 
self of every opportunity of becoming familiar with its action. 


The student having been thus placed in possession of the 
more valuable methods for preparing the brain for the 
imbedding microtome, it will be well to indicate the method 
he should adopt in his earlier attempts, as each process has its 
own individual merits. It is advisable that his first attempts 
should be made with comparatively small portions of tissue 
— about 3 c.c. in bulk — and that he should employ the 
process of hardening by Miiller's fluid and potassium bichro- 

1. This process, to which I assign the first place amongst 
the chrome-hardening processes, is slow but very certain. The 
results are, to my mind, more satisfactory than those of any 
of the other methods, and I invariably adopt it myself as the 
one for general use at the West Riding Asylum. The vessels 
which the student will find most convenient to use for harden- 
ing these smaller portions of tissues are the stoppered glass 
bottles of 4-oz. capacity employed for dispensing purposes. 
Each bottle should be labelled with the name of specimen and 
date and nature of each successive change of reagent, whilst 
the specimen should occasionally be removed, and its con- 
sistence noted so as to familiarize the touch with the increas- 
ing firmness of the tissue, and the degree of hardness 
requisite. The method recommended by Rutherford (2) is 
also very reliable for moderate-size specimens. The methods 
advocated by Betz and Hamilton (3, 4) are peculiarly well 
adapted for large segments of the brain, a whole hemisphere 


being thus readily hardened throughout. When these large 
masses of tissue have to be dealt with, special precautions are 
requisite. Thus the hemisphere should be sliced horizontally 
or vertically into segments not over f-inch in thickness. 
Each segment should rest upon a bed of cotton-wool in the 
vessel for its reception ; and if two segments are included, 
another stratum of cotton-wool should be interposed betwixt 
them. The vessels containing the specimen should be of 
large size, varying in capacity with the bulk of the latter. 
Thus a large preparation glass, such as is used for preserving 
brain in spirit on museum shelves, mil be found well adapted 
for hardening the whole hemisphere ; whilst pickle-bottles, 
esjDccially those provided vdth. the " patent lever stopper," are 
most suitable for the basal ganglia, cerebellimi, smaller por- 
tions of the cerebrimi and the pons. Still smaller segments, 
such as the student will have more frequently to deal with, 
are best hardened in the 4-oz. stoppered bottle, a good 
supply of which should be kept on hand. All the foregoing 
processes are contrasted with that of Exner's (osmic acid) 
in being most adapted for the demonstration of the nerve- 
cells and plexuses of the grey matter of the brain and its 
ganglia ; whilst the latter, failing in this respect, is, on the 
other hand, infinitely better suited for exhibiting the struc- 
ture of the cerebral medulla and its extensions into the cortex 
of the brain. The student is therefore recommended to make 
himself acquainted with the structures of the cortex by the 
chrome methods, and subsequently to employ Exner's method 
for demonstrating the medullated tracts of the cortex. 


Having satisfied himself that the specimen is sufiiciently 
hard for section-cutting, the student first cuts off a portion 
which can be accommodated by the well of the microtome, 
and prepares it for imbedding by a prior immersion for a 
few minutes in rectified spirits. This removes the fluid 
derived from the chrome solution from which it has been 


taken. In the meantime the imbedding mass should he 
melted at the lowest temperatm-e requisite. The micrometer- 
screw should he lowered sufficiently to allow of a deep imbed- 
ding of the tissue. The preparation is next removed from the 
spirit, rapidly dried by a fold of blotting-paper, and momen- 
tarily plunged into the warm imbedding mixtiu'e, which, upon 
its removal, leaves a film over its surface, whilst all small 
cavities or lacuna which it may contain are filled up. The 
melted mass is now poured into the well of the microtome, 
and the preparation immersed and held in the position re- 
quired for section-cutting until fixed by the cooled and con- 
solidated mass. If the imbedding mass is one subject to 
much contraction on cooling, it is requisite, just as it is 
becoming " set," to press down with the end of a spatula the 
margin of the wax and the oil mass against the sides of the 

The Imbedding Mass. — In the process of imbedding, 
the principle to be remembered is that the mass should be not 
too resistant to the blade, whilst at the same time it affords 
efficient support for the imbedded tissue. If the textm-e be 
one readily permeated by the imbedding mixture, contraction 
of the latter in cooling is apt to result in injmy to the prepa- 
ration, more especially if this be one of the more dehcate 
structures. Thus, in the delicate embryos of the fowl and 
other similar organisms, special methods of imbedding will 
be requisite. For the brain and spinal cord, however, the 
process is comparatively simple, since the imbedding mass 
neither permeates its structures nor injures it by contracting. 

The following are the more important media employed for 
imbedding prior to section-cutting : — 

1 . White wax. Olive oil, equal parts. 

2. White wax, 3 parts. Olive oil, 1 part. 

3. Wliite wax. Cacao -butter. 

4. Solid paraffin, 5 parts. Hog's lard, 1 part. 

6. Solid paraffin, 5 parts. Paraffin oil, 1 pai-t. Hog's lard, 1 part. 

6. Solid paraffin, 5 parts. Spermaceti, 2 parts. Hog's lard, 1 part. 

7. Spermaceti, 4 parts. Cacao-butter, 1 part. 

8. Spermaceti, 4 parts. Castor oil, 1 part. 


The first on this list is perhaps the more generally employed 
medimn, but it possesses the disadvantage of contracting 
strongly on cooling, so as to leave an interval betwixt the 
wax plug and the sides of the well : this, of course, will 
necessarily tend to loosen the mass, and allow rotatory or 
" wobbhng " movements, which prevent accurate section- 
cutting. These disadvantages may be overcome by one or 
other of the following measures : — 

Yary the proportion of the oil and wax in favour of the 
former. When solidifying, press down the edge of 
the mass with a spatula. Use a microtome plug wdth 
groove or projections on upper surface. Drive the 
mass out of the well, partly surround it with a narrow 
strip of blotting-paper, and return it — forcibly pressing 
it down into the well — the blotting-paper swells by 
imbibition of the spirit in section-cutting, and so fixes 
the plug firmly. 

In the use of the third medium, viz., white wax and cacao- 
butter, far less contraction ensues upon solidification, and a 
firm supporting mass is obtained, the proportion of the in- 
gredients varj'ing with the firmness required. I can also 
speak favourably of the combination of paraffin with hog's 
lard. In all these cases it is well to keep the mass in a 
small tinned pot with lip, provided with cover and handle, so 
that it can readily be held over a gas-jet or Bunsen burner, 
and melted as required. Another imbedding agent used is 
gum, which is solidified by the application of alcohol or 
methylated spuits. This method, proposed by Briicke, 
although valuable for many tissues, cannot be recommended 
for brain. ^ 

Imbedding is wholly dispensed with in the form of micro- 
tome devised by Schieif erdecker, where the preparation having 
been hardened by alcohol as far as practicable, is placed in 
the well and clamped firmly by the arrangement already de- 
scribed. In the small hand-microtome of Eanvier, again, the 

1 " Handbook of the Physiological Lahoratoiy," p. 92. 


ordinary melted media for imbedding are often exclianged 
for elder-pith, whicli is packed around the tissue in the well 
in a dry state, and then, upon immersion in spirit, the pith 
swells and firmly fixes the preparation. None of these latter 
methods are so suitable for hardened brain as imbedding in 
the wax and paraffin mixtures. 

Section-Cutting. — The preparation being satisfactorily 
imbedded, and the mass perfectly cold and hard, our next 
procedure is to cut the finest possible sections — they cannot 
he cut too fine. Place on the table supporting the microtome, 
and in front of the latter, two vessels, one a Griffin beaker- 
glass of three inches diameter, and the other a cylindrical 
glass jar, a brain-preparation jar, or better still, a flat-bottomed 
porcelain evaporating basin. Nearly fill the beaker with 
methylated spirit, and the latter with water. Seat yourself 
in front of the microtome at a convenient level for the free 
play of the hands, place a soft towel over the knees, to be 
used for wiping off adherent wax from the imbedding mass. 
Have close at hand, also, a razor-strop, a porcelain capsule of 
small size, and some camel-hair brushes. 

Dip the section-blade in the methylated spirit, and having 
raised the imbedded mass slightly above the level of the sec- 
tion-plate, slice off the superficial portion, exposing a clean 
smooth siu'f ace of the tissue for cutting. Again dip the blade 
into the spirit, and turning the micrometer-screw, say through 
one-eighth of a revolution, place the flat of the blade upon the 
section-plate in front of the mass, with the back towards 
yourself and the handle in your right hand, grasped near the 
heel of the blade. Now cut your first section by a clean 
steady sweep from heel to point, and always aicay from your- 
self. No saw-like movement should be performed, nor should 
the blade be arrested occasionally in its course, the section 
shaved continuously and evenly off the preparation. The 
thin film of wax removed from the front of the tissue is 
sometimes likely to get in the way of very fine sections, and 
hence it is usually advisable, with a penknife to gently 


remove all the imbedding mass in front of the preparation 
to about the depth of a couple of millimeters prior to section- 
entting. Subsequent sections should be cut with a still more 
restricted movement of the screw, until the operator has foimd 
the minimum degree of movement at which a section can be 
cut. This, of coiu-se, is a matter of tact, and the student 
will find that he daily improves the quality of his sections 
by practical manipulation. Two precautions must ever be 
taken if successful sections are desii-ed. The first is, that as 
the blade is raised out of the spirit, the latter should not be 
drained off, but a good flow be retained over the surface of 
the blade, so that the section is floated up as it is cut. In 
the next place make it a rule to pass the blade across the 
strop frequently, after cutting half-a-dozen sections, as it 
requu-es to be kept at the flnest possible edge. As the 
sections are cut they are floated off into the porcelain basin 
of water, and are freed from adherent particles of the 
imbedding mass by the rapid gyratory movements they here 
tmdergo from the euiTents caused by the attraction of water 
for the spirit they contain. After floating off each section 
in the water, pass the blade softly over the towel to clear its 
surface from adherent wax, prior to bathing it afresh in the 
spiiit. How is the student to ascertain whether his sections 
be sufficiently thin ? The exact thickness of the section is 
ascertained if the pitch of the micrometer-screw is known, and 
the degree of movement which he adopts noted. Thus, if the 
screw have eighty threads to the inch, a movement through 
one-eighth of a complete tm-n, i.e., 45°, will give a section 
■g-J-Q-inch in thickness, whilst a movement through an 
arc of 30°, or one-twelfth the entire turn, gives a section 
g-^-o-inch thick. He will, however, be not far fi'om wrong 
by following this rule : reject all sections which do not float, 
and of the latter preserve as valuable only those which appear 
as a mere film on the surface of the fluid, requiring to be 
looked at sideways to be distinctly seen. It is at this stage 
of the proceeding, that of section-cutting, that we can best 
judge of the merits of the imbedding mass, which, if found 


unnecessarily resistant to the blade, should be modified by 
the addition of more of the diluting adjuncts, viz., lard, oil, 
or cacao-butter. 

The Section Blade. — This is an item of supreme im- 
portance to the histologist, and it is by no means true, as we 
often find asserted, that any razor blade answers the pm^pose 
equally well with the specially prepared knives. I have 
found this statement made frequently by those who were by 
no means fortunate in the thinness and delicacy of their 
sections ; whilst all with whom I am acquainted, who can 
rejoice in the good qualities of their sections, have always 
paid great attention to the section-blade which they employ. 
The qualities of a good blade for section-cutting upon the 
microtome depend upon its make, form, edge, and tempering. 
It shoidd be sufficiently long to ensure a free sweep over the 
whole surface of the section-plate from heel to point, wide 
enough from back to edge to support and float up the 
largest-sized section which might have to be cut ; it should 
be hollowed out on both sm-faces, but be most concave on the 
sui'face uppermost in section-cutting ; the edge should be 
extremely sharp, and perfect, whilst the angle formed by the 
converging sto-faces here should not be so acute as to involve 
bending of the blade when subjected to slight pressure ; the 
quality and tempering of the steel should be of the best 
character. The above points are all most essential require- 
ments in a good section-blade ; but another important con- 
sideration is that the plane of the back and edge of the knife 
should exactly coincide when it rests upon a perfectly level 
surface as the microtome section-plate — not the slightest 
degree of tilting being admissible. 

For the smaller microtomes I have invariably used razor- 
blades, my three instruments measuring in length and width 
of cutting- surface as follows : — 

No. 1 . 4 inches x 1 incli (made by Weeclon) . 



X 1 ,, 






X U M 





The first is a most valuable knife. It is fitted into a short, 
firm, and fixed handle. The latter, which is my favourite 
blade, has a folding handle, and is sketched in outline with a 
section of its concave surfaces in an early number of Brain} 
Some of the earliest and most discouraging failures upon the 
part of the tyro in section- cutting depend upon the condition 
of the blade, and it is necessary that he should be fully 
impressed with the fact that not only must he secui"e a couple 
of section-knives with the above qualities, but that he should 
jealously keep them in most perfect order. To ensure 
the latter condition, let him observe the following precau- 
tions : — 

Use no undue pressm^e against the section-plate whilst cut- 
ting, and keep the surface of the blade absolutely level with 
the plate. Pass it frequently along the strop whilst cutting 
sections. After each day's work examine its edge most 
critically, and if the slightest irregularity or notching be 
apparent, it must be reset before use. 

Keep the blade dry and well polished by a soft handker- 
chief, and if in a closed handle, have a chamois-leather case 
made for it. - 

Subsequent Treatment of Sections. — The sections 
have now to be removed carefully by a camel-hair brush to 
methylated spirit contained in the small porcelain capsule, in 
which they may be allowed to soak for some time to remove 
all traces of chrome, which otherwise interferes with the 
subsequent staining they have to undergo. 

Large Sections through Hemisphere. — The above 
account refers especially to the smaller class of sections. 
When it is requisite to obtain sections upon a much larger 
scale, such as those thi'ough the hemisphere or the whole brain, 
the process requires modification. The method of imbedding 
is precisely similar in all its details, but when we come to 

^ V'nh- Brain, part iii. p. 3r>3. October, 1878. 


cutting the section, special forms of knife are found requisite. 
The blade which I use measures 16 inches long by 2 inches 
wide, and is fixed in upright handles at each end. The zinc 
basin in which the large microtome rests is filled with water, 
and then sections are cut with a saw-like movement of the 
blade, and not, as in the former cases, by one complete sweep 
through the structures. It requires a steady hand and much 
manipulative skill to obtain these larger sections both thin 
and perfect, and the student will only perfect himself by 
frequent and persevering efforts. These large sections can 
be readily removed by means of sheets of paper upon Avhich 
they are floated, and this support may be given to them 
throughout the various subsequent stages of preparation. 
When the desired number of sections have been obtained, 
the imbedded plug may be removed, the remaining portion 
of tissue placed in a 0*5 per cent, solution of bichromate of 
potash, until required. The microtome should be carefully 
dried and placed away in a drawer for safety, as the slightest 
scratching of the polished section-plate will ruin a good blade 
in a few minutes. 

Apparati;s for Hardening Tissues and 

Winchester bottles for hardening reagents. 

Jars and 4-oz. phials for hardening tissues. 

An ice safe. 

Microtome for imbedding. 

Microtome for freezing. 

Griass " preparation- j ar " for receiving sections. 

Imbedding mass. 

Spirit lamp. 

Porcelain capsule. 

Griass beaker for methylated spirit. 

Ether spray apparatus. 




The staining reagents advocated by different authorities 
for sections of the nervous system prepared by hardening are 
numerous, and the list has become so lengthened since the 
earlier labours of Lockhart Clarke, Van-der-Kolk, and others, 
that one great difficulty presented to the student is to make 
a judicious selection of such as will yield him the best results 
in the special direction pursued by his work. Amongst 
those more generally used are carmine and its combinations, 
picro-carmine, indigo-carmine, and borax- carmine ; the 
aniline series, comprising aniline blue-black, auiline blue, 
methyl-aniline, rosanilin or magenta ; hoematoxylin or log- 
wood ; eosin ; picric acid ; osmic acid ; double chloride of gold 
and potassium. Many years' experience has convinced me 
that a large proportion of these reagents may be safely dis- 
pensed with. The following really comprises all essential 
staining solutions for nervous tissues : — 


Carmine in ammoniacal solution. 


Aniline blue-black. 

Aniline blue. 

Osmic acid. 

The only solution comprising a double pigment in this 
list is the picro-carmine, but double staining may be exten- 
sively employed by the combined agency of the above and 
two other dyes, the more useful being as follows : — 

List of Pigments for Dourle Staixtxo. 

Aniline picro-carminate. 


Osmium with picro-cannine. 

Hoematoxylin with anilin. 

Hsematoxylin with eosin. 

Picro-carmine witli iodine green. 

I 2 


In making clioice of metallic impregnation and reduction 
by osmium or gold, or of staining by the mineral or vegetable 
pigments in the list given, the student must be influenced by 
several considerations. 

Does he require a uniform staining by a simple dye, or 
does he wish for differentiation of elements by the use of 
double pigments — the process of double staining ? Does he 
wish to examine more specially the cortex or medulla of 
the cerebrum, or the cortex of the cerebellum, or again, 
the pons, medulla oblongata, or spinal cord ? 

Does he desii-e to display certain particular constituents of 
the cortex — e.g.^ the nerve-cells and their processes — to the 
fullest possible extent, even at the sacrifice of other structures, 
such as is often required in minute investigations into the 
histological constitution of tissues ? 

Whatever be his object, these questions must receive an 
answer before the staining reagent can be selected which will 
yield him the effect desired. I propose to give here the com- 
position of each reagent, the methods of staining, the special 
value of each dye for the various histological elements, and 
to supplement these observations by an analysis of their 
relative merit for different regions of the cerebro-spinal 


1. The Dye. — This reagent has often fallen into great 
disrepute, owing to the notable variability in the quality 
of the solutions issued by different makers. The student is 
strongly recommended to make his own solution, adopting 
the formula given by Minot or that employed by Kleinen- 
berg. For the brain and spinal cord I use the former, as 
preferable to that of Kleinenberg, and I find it at all times 
most trustworthy in its reaction, and in the uniformity' of 
results obtained. 

prepakatio^ of hardened brain. 117 

Minot's Formula. 

Hsematoxylin (crystals) ... ... 3"5 parts. 

Absolute alcohol ... ... ... 100"0 ,, 

Alum ... ... ... ... ... I'O ,, 

Water 300-0 „ 

First dissolve the htematoxylin in the alcohol, and add to 
it the alum, previously dissolved in the water. Keep in a 
closely-stoppered glass bottle labelled " hsematoxylin dye 
(Minot)." For use, a solution of alimi (0*5 per cent.) is 
poured into a watch-glass or porcelain capsule, according to 
the size and number of sections to be stained. A little of 
the dye is then dropped in, until the solution assumes a light 
violet tint, when it should be carefully filtered before the 
sections are immersed. The degree of dilution will soon be 
learnt after a little practice. 

Kleinenberg's Formula. 

a. Make a satm-ated solution of crystallized calcium chloride 

in alcohol (70 per cent.), and add alum to saturation. 
h. Make a satm-ated solution of alum in alcohol (70 per cent.), 

and add Solution 1 to Solution 2 in the proportion of 

1 to 8. 
c. To the resultant mixture now add a fetv drops of a barely 

alkaline saturated solution of hsematoxylin. 

Excellent as this logwood solution is for embryonic tissues, 
I cannot recommend it for the sections of hardened brain, 
such as we are now engaged with, to the same extent that I 
can Minot's solution. 

2. The Staining Process.— Withdi'aw the sections to 
be stained from the methylated spirits in Avhich they float 
by means of a glass rod or camel-hair pencil, and immerse 
them in the logwood dye contained in a watch-glass or porce- 
lain capsule. The sections should be lightly stained, other- 
wise they are spoilt by too diffuse a ooloming and by 


acquiring a brittleness unfavourable to subsequent manipula- 
tion. In his first attempts the student should examine a 
section occasionally on the stage of the microscope, and so 
learn to judge of the progress made by the staining. When 
sufficiently stained, the dye should be poured off, the sec- 
tions floated up in a porcelain capside half full of water, the 
capside slightly inclined towards a gentle stream of water from 
a tap which, falling upon the edge of the vessel, keeps up a 
constant change of fluid and ensiu'es a most thorough washing 
of the sections in the currents it produces. Care must be 
taken that the delicate sections are not torn by this means, 
which is really necessary to ensure removal of the deposit 
which adheres to their surface. Next remove the sections to 
a capside containing methylated spirit, and in one or two 
hours all tendency to diffuse staining will have disaj^peared. 
They are now placed for five or ten minutes in rectified 
spirits or absolute alcohol for dehydration, transferred to the 
centre of a glass slide, floated up by a drop or two of oil of 
cloves, which, when it has permeated the tissue and rendered 
it perfectly transparent, is drained off, and the section 
mounted in Canada balsam. 

To ensui'e successful staining with this reagent, the follow- 
ing points must be carefully attended to : — 

Stain only the very finest sections, 

Ijet the sections be perfectly freed by spirit from any 
acidity (chromic or picric acid, etc.), otherwise the staining 
will be a failure. 

Make your own solution of hcematoxylin. 

Employ Minot's solution in preference to others, and 
reserve Kleinenberg's for embryonic brain. 

Never omit the proper dilution and filtering prior to use. 

Guard against diffuse staining and brittleness from too 
prolonged action of the dye. 

Wash the sections very tJiovoiigJiIy subsequent to staining. 

Remove any accidental diffuse coloration by long im- 
mersion in spirit. 


Dehydrate perfectly before clearing up with, clove oil. 
Mount in a benzole solution of balsam. 

3. Notes on the Reaction of the Dye. — Hsema- 

toxylin is specially well suited for the display of the various 
nuclei met with in brain and spinal cord, — e.g., the con- 
nective nuclei of neuroglia, the peri-cellular and peri- 
vascular nuclei, the nuclei and nucleoli of the nerve-cells. 
The larger nerve-cells of the cortex are beautifidly shown in 
successful preparations, and their primary and secondary 
branches may be followed for some distance, but never to the 
extent seen in aniline staining. On the other hand, both 
neuroglia basis and nerve-structm-es undergo much shrinking 
by the use of the dye, and from this cause the smaller cells 
of the upper layers lose, to a great extent, their normal 
features. The protoplasmic extensions and nerve-fibre net- 
work of the cortex are indeed very poorly exhibited in 
logwood, as compared with aniline preparations. The axial 
cylinders of medullated fibres exposed in transverse section 
are well exhibited by logwood, and hence the sections of 
spinal cord and medulla, stained by the dye, show to 

Carmine Staining. 

1. The Dye. — The most useful solution for hardened 
brain sections is that recommended by Beale, diluted to the 
required strength, the original solution being too strong for 
our piu-pose. 

Beale's Formula. 

Carmine (in small fragments) 

10 grains. 

Strong solution of ammonia 

1 drachm 

Price's glycerine 

2 ounces. 


h „ 

Distilled water 

2 „ 

Agitate the carmine with the liquor ammonige in a test tube 
gently heated over a spirit lamp. When solution is complete, 


carefully boil for a few minutes and expose tlie open test tube 
for an hour to allow excess of ammonia to escape. Add the 
water, filter, and after the addition of the spirit and glycerine, 
expose to the air until the odour of ammonia given off from 
it is very faint. Keep the clear fluid in a stoppered bottle, 
and whenever carmine becomes precipitated, add a drop or 
two of liquor ammonitB. 

Formula for Borax Carmine. 

Carmine ... ... ... ... 1 part. 

Borax ... ... ... ... ... 4 ,, 

Water ... ... ... ... ... 56 „ 

Alcohol q.s. 

Dissolve the borax in the water, and add the carmine. Filter 
the solution, and add just sufficient alcohol to ensure free 
permeation of the tissue by the dye. 

Thiersch's Formula. 

Solution A. Carmine ... ... 1 part. 

Liquor ammonifc ... 1 ,, 

Distilled water ... ... 3 , , 

Dissolve and filter. 

Solution B. Oxalic acid. ... ... 1 part. 

Distilled water ... ... 22 „ 

Mix one part of Solution A with eight parts of Solution B, 
and add twelve parts of absolute alcohol. Modifications of 
Grerlach's original carmine dye have also been introduced by 
Frey and others, but the three above given are all that are 
really requisite for staining nervous tissues. 

2. The Staining Process. — Give the preference to 
Beale's solution, although its density from the glycerine 
renders the action slow. Dilute the strong solution with 
seven times its bulk of water and filter. The sections should 
be placed in a comparatively Jorge quantity of the dilute 


solution covered from dust and left to undergo very gradual 
staining, a process wliicli may occupy eight hours, or even 
longer. "When a section removed to the microscope stage 
shows sufficient staining, the reagent may be poured off, 
sujjerfluous carmine removed by gentle washing, and all the 
sections immersed in a dilute solution of glacial acetic acid 
(0"5 per cent.) for fifteen to twenty minutes. The acid 
deprives the specimen of all diffuse staining, and fixes the 
dye more especially in the germinal centres, whilst at the 
same time the tint is brightened by its agency. Great care 
should be taken that neither section nor staining solution is 
very alkaline in reaction, otherwise the staining will prove too 
deep and diffuse, and the structure itself injuriously affected. 
A minimum amount of ammonia, however, must be present 
in the dye just recognizable by its odour, and if this be not the 
case, a drop or two of the liquor ammonite should be added 
to Beale's strong dye prior to making the dilute solution. 
These sections may now be washed and mounted in glycerine 
or Farrant's solution, or they may be dehydrated in the usual 
way by spirit, cleared up in oil of cloves, and mounted in 
Canada balsam or dammar. 

With a few of these carmine-stained sections the student 
shoidd try the following modified process of mounting, 
whereby valuable information may be obtained. Several 
years ago I described a method of displaying a great wealth 
of stmctiu'e on the cortex cerebri by altering the refractive 
indices of the structm-al elements, and so producing very 
remarkable differentiation of structui'e.^ I there stated what 
follows : — " On placing an unstained section of cerebrimi or 
cerebellum satiu*ated with spirit in the field of the micro- 
scope, little or no structure is apparent, but if a drop of 
essential oil be now allowed to run over it, there will be 
observed at a certain stage of clearing up, and whilst the spirit 
is evaporating, a sudden starting out in bold relief of the 
cells, nerve-fibres, blood-vessels, etc., which again disappear or 
partially fade on perfect clearing of the section. Now, this 

' Quarterly Journal of Micros. Sac. vol. xvi. 


appearance may be fixed by suddenly dropping over its 
sm-face a little balsam and permanently mounting. Upon 
this fact depends the process now to be described. Sections 
treated with Beale's carmine solution (1 to 7) and washed with 
the acid, are placed, saturated with spii'it, upon a slide. 
When the spirit has nearly all evaporated, a drop of oil of 
anise is allowed to flow over the section (not to float it up), 
and the clearing is watched on the stage of the microscope ; 
then, just when the appearance referred to above is presented 
to view, a drop of balsam is allowed to run over the section, 
and a covering glass permanently fixed on. In lieu of oil 
of anise, I frequently employed glycerine with the same 
results, mounting the specimen in glycerine jelly. * * * 
I ^vill state my belief that this method will yet prove of most 
essential service in the estimation of the relative proportion 
of cell-processes in any individual section, and the most 
accurate tracing of any existing connections, for not by the 
deepest aniline staining have I yet succeeded in demonstrating 
the existence of so thick and numerous a series of j)rocesses 
diverging from the pyramidal layers of the cerebral cortex 
as by the method described above." It is satisfactory to find 
these opinions confirmed by Prof. Stirling in his recent text- 
book of " Practical Histology." Dr. Stirling says : — " The 
processes of the cells (of the cerebrum) are best seen in pre- 
parations which are only iMrtialhj cleared up imder the 
influence of clove oil. This is a most important method of 
investigation. I have often seen in this way delicate fibrils, 
not unlike elastic fibres, and which are not distinct when 
the section is completely cleared up."^ 

3. Notes on the Reaction of the Dye. — Carmine is 
especially suited as a dye for the large nerve-cells, their 
contents, nuclei, and nucleoli. It exhibits well the connective 
cells and the vascular apparatus. For the large ganglionic 
cells and their immediate environment, nothing can perhaps 
sm-pass a successful carmine preparation, whilst it does not 

» "Text-book of Practical Histology," p. 101. Smith, Elder, & Co. 


appear to produce any further slirinldng of protoplasm or 
connective such as is produced by hsematoxylin. It is less 
adapted for displaying the cell-processes, and wholly fails to 
exhibit the details of structure in smaller nerve-cells, such as 
those of the upper cortical layers. It has been regarded, 
even by so eminent an authority as Charcot, as the only 
reliable dye for exhibiting the morbid state regarded as 
sclerosis of the different columns of the cord. This, how- 
ever, is not the case, since aniline blue-black poi'traj's the 
lesion even more distinctly. The more important objections 
attached to this dye are the variable results given ; the little 
definition often afforded of the tissue elements from the 
tendency to diffuse staining ; the unpleasant glare of the 
ca]*mine tint for continuous work, and its unsuitability for 
examination by artificial light. 

Picro-Carmine Staining. 

1 . The Dye. — This valuable reagent, introduced by 
Hanvier, is of very special use in the preparation of nervous 
tissues. It is readily made as follows : — 

Ranvier's Formula, 

Best carmine ... ... ... 1 gramme. 

Water ... ... 10 c.c. 

Liquor ammonise ... ... ... 3 c.c. 

liub the carmine up with water, add the solution of 
ammonia to the fluid in a test tube, and aid the solution of 
the carmine by gentle heat carefully a2:)plied. When dis- 
solved and the solution perfectly cold, pour it into 200 c.c. 
of a saturated aqueous solution of picric acid. Place the 
solution in an open vessel and Tvith gentle warmth evajiorate 
to ono-third of its bulk. Filter and keep in a stoppered 
bottle labelled " Ranvier's picro-carminate dye." 

Picro- carmine may be purchased in a solid form as a 
granular and imperfectly crystalline substance — a 1 per 


cent., solution is then used. From personal experience I 
cannot commend its use, and I would strongly recommend 
the preparation after the formula above given. 

2. The Staining Process. — Immerse the sections in a 
small quantity of the picro-carminate solution for a period of 
from twenty to thirty minutes. Pour off the dye, draining away 
all the superfluous fluid, and then, without urishing the sectiom, 
float them up by glycerine in which they must be mounted. 
It will often be found advisable to stain each section upon 
the slide, dropping over sufficient of the reagent to cover it — 
to clear it up with glycerine and mount it permanently upon 
the same slide. Picric acid, being soluble in water, is readily 
removed upon washing these sections ; hence, to preserve the 
colour due to both pigments, we must carefully avoid washing 
with water.^ 

Upon the other hand, washing freely, dehydrating with 
spirit, and mounting in the usual way in balsam, will yield 
us excellent carmine preparations, and so far as my 
experience goes, better specimens than those subjected to 
the action of the simple ammoniacal or borax solutions. 

3. Notes on the Reaction of the Dye. — Picro- 
carmiiie has the special merit of showing less tendency to 
diffuse staining than simple carmine dyes. The presence 
of the two pigments appear notably to restrict their action 
to the germinal centres, for which they each have affinity. 
Elastic tissue and muscle will be stained yellow, whilst nuclei 
and connective take the carmine tint. Minute nervous 
plexuses are poorly shown, as they occur in the structure of 
the cortex ; hence, the nerve-cell processes and their ramifi- 
cations can rarely be followed to any distance. The structure 
of the investing membranes and the vascular apparatus is 
better defined by this reagent than any other we are acquainted 
with. In fact, the merits and demerits of carmine staining, 
pertain to the picro-carminate, although, for powers of 

1 This difficulty is also overcome by washing in a saturated aqueous solu- 
tion of picric acid, and dehydrating in an alcoholic solution of the same 


differentiation, it is superior to simple amraoniacal carmine, 
and its roJe as a staining- reagent more extensive. For com- 
bination with other dyes, such as aniline, logwood, and osmic 
acid, it is peculiarly useful. As a general dye for nerve- 
structures, it ranks next to aniline and aniline picro-carminate, 
and is far superior to logwood. 

Aniline Staining. 

1. The Blue-black Dye.— This, which is by far the 
most valuable of the aniline series of dyes for the brain and 
spinal cord, is thus made : — 

Aniline blue-black 1 gramme. 

Distilled water 400 c.c. 

Dissolve, filter, and keep in a stoppered bottle, labelled 
" anihne black dye, aqueous solution, 0-25 per cent." It may, 
however, be convenient to keep a stock solution of 1 per cent, 
strength, as rapid staining is occasionally required, and to 
dilute, as required, to 0-2-3 or O'o per cent., always filtering 
these dilutions prior to use. The solution, as above given, 
may be employed for staining fresh and hardened sections, 
the aqueous solution being the best dye we possess for the 
former. The alcoholic solution, as first recommended by Mr. 
Sankey, was the following : — 

Aniline blue-black ... 5 centigrammes. 

Water ... ... ... 2 cubic centimeters. 

Dissolve and pom- into it 99 c.c. of methylated spirit. 
Filter the solution, and label "alcoholic solution of aniline 
black, O'Oo per cent." 

Professor Stirling advises the use of a solution double the 
strength of the above, thus : — 

Aniline blue-black ... 1 decigramme. 

Water ... ... ... 4 cubic centimeters. 

Dissolve, and add 100 c.c. of rectified spirit, and filter. 
Label, " anihne blue black, alcoholic solution, O'l per cent." 
I much prefer the aqueous solution for most pm-poses. 


2. The Staining Process. — Sections of liardened cortex, 
pons, medulla, and spinal cord, may alike be left in the 
aqueous solution (0*25 per cent.) for one hour. In this 
time they will, generally, be found stained to a sufficient 
extent, when they must be removed to a vessel containing 
water, and well washed from superfluous dye. Next, dehydrate 
by rectified spirit or absolute alcohol ; clear up by oil of 
cloves, and mount in balsam or dammar. 

Sections of Cerebellar Cortex shoidd be deeply stained 
by aniline, gently washed in water, and immersed for 
twenty to thirty minutes in a solution of chloral hydrate 
(2 per cent.). Next transfer them to the following solu- 
tion : — 

Solution of chloral (2 per cent.). 

Oil of cloves equal parts. 

Alcohol ... ... ... q. s. to dissolve per- 

fectly, and form a clear solution. Add the alcohol by 
degrees, stirring the solution with a glass rod, and avoiding 
excess of spirit. During use, carefully cover the watch- 
glass containing this solution, so as to avoid evaporation. The 
chloral removes diffuse staining, whilst the clove oil clears up 
the section, and enables us, by examining one occasionally 
under a low magnifying power, to decide when they have 
reached a satisfactory stage. When this has been attained, 
remove your section to a slide ; rapidly wash with a little 
pm-e alcohol ; thoroughly clear with clove oil, and mount with 
balsam. The alcoholic solution of aniline also stains sections 
of cerebrum, pons, and medulla, and must be cleared and 
mounted in the usual Avaj^ Grlj^cerine being a powerful 
solvent of aniline, must not be made the medium for mounting 
these sections. 

3. Notes on the Reaction of the Dye. — Its action is 
energetic, and its results are certain and constant. It is 
readily controlled, so that any depth of staining may be 
obtained with ease. The tint given varies from a bluisli-grey to 


a deep bkie-'black, and is a pleasant one for the eye, causing 
little or no fatigue. It enables us to obtain the clearest and 
sharpest definition of elements in a tissue, without modifying 
their structm-e by shrinking or other change. No otlier 
staining reagent displays the nerve-cell processes to sucli a 
remarkable extent as does this dye, the very finest ramifica- 
tion being followed out with ease. It has a special affinity 
for protoplasm, staining the nuclei of the nerve-cells most 
deeply, then the nerve-cells themselves and their protoplasmic 
extensions, and, to a less intense degree, the nuclei of 
the neuroglia and perivascular walls. The finely-formed 
nerve and connective mesh-work, forming the basis substance 
of the cortex, is stained of a pale grey. This dye fails to 
produce any action upon the medullated sheath of nerve- 
fibres, affecting only the axis-cylinder. Transverse sections, 
therefore, of nerve-fibres, such as are seen in cross sections of 
the medulla, cord, and nerve-trunks, exhibit the axis-cylinder 
stained of a bluish-black tint, the unstained white matter of 
Schwann siuTOunding it ; whilst the intertubular connective, 
lightly stained, differentiates the medullated tubes from each 
other. The value of this clj^e for connective tissue is very 
limited, its role being chiefly confined to the nerve-elements. 

OsMic Acid Staining. 

This method we owe to Professor Exner. It is peculiar 
in the fact that the hardening and staining proceed 
simultaneously, the same reagent being employed for both 
purposes. The preparation of the hardened brain hy im- 
mersion in osmic acid has ah-eady been alluded to, and we 
have now simply to detail his further treatment of the 
sections so obtained by the microtomo. The sections, owing 
to the deep staining they undergo, must be extremely thJn 
and should be placed in glycerine immediately, since pro- 
longed immersion in alcohol injm-es them seriously. Transfer 
a section to a glass slide and add a few drops of strong liquor 
ammonia?. Absorb all superfluous moistiu'e around the section 


with blotting jiaper, and after waiting a short time for the 
ammonia to effect the section thoroughly, place the cover 
glass on without injury to the preparation which has heen 
softened by the reagent, and examine immediately on the 
microscope stage. It is at this early period that the section 
shows to greatest advantage. For permanent preservation 
Exner surrounds the cover glass with the soluble silicate 
called water glass. Formerly an ammoniacal carmine solu- 
tion was used, but Exner found that the results obtained 
could be attributed only to the reaction of ammonia, and so 
he has dispensed with any carmine staining as the osmium 
suffices for all pui-poses. 

Notes on the Value of this Method. — The meduUated 
fibres can be traced upwards into the highest realms of the 
cortex, exhibiting a wealth of structure which no other 
method displays. The meduUated sheath is deeply stained 
and can be traced distinctly in the case of the smallest nerve- 
fibres. The nerve-cells, although stained, do not form such 
prominent objects as by other methods of staining ; this 
process being especially adapted for tracing the ultimate 
course of meduUated fibres through the difPerent layers of the 
cortex and determining their final destination. Exner states 
that the staining of the minute fibres occasionally disappears 
in time, and the student should also be aware that the brain- 
structm'e swells up so much under the agency of ammonia 
that the section is increased by one-third its full diameter.^ 

On the Use of Compound Pigments. 

Under this head I would include some of the most valu- 
able methods we possess for delineating the minute structures 
of the nervous system. Picro-carmine, as consisting of two 
pigments, would naturally fall under this category, but since 
it is employed in the same way as the simple dyes, I have 

^ For results obtained by Prof. Exner with this method, sec Original 
Memoir (op. cit.), or abstract of the same in Brain, part xv. 


found it more convenient to include it amongst them. The 
methods of staining to which I now refer are those in which 
the tissue is first submitted to one dye, and the result subse- 
quently modified by the addition of a second pigment. In 
these cases, either different histological elements assume the 
tint of each pigment, or the former staining is greatly 
improved in character by the second pigment combining and 
modifying the tint acquired. In the former case only does 
a genuine double staining occur ; in tlie latter we obtain a 
single tint, but one of valuable quality. 

1. Aniline Picro-carminate Staining. — Three pig- 
ments are here used, viz., the aqueous solution of aniline 
blue-black (0"25 per cent.), and Eanvier's compound dye, 
picro-carmine. Sections, which should be very thin, may be 
stained on a glass slide, by dropping over them from a pipette 
sufficient picro-carmine to cover the section completely. 
When deeply stained by this reagent the latter is di'ained off 
and the section covered in a like manner by a quantity of the 
aniline blue-black solution. It will be found that the action 
of the aniline proceeds much more rapidly upon a section 
stained by picro-carmine — ten minutes usually sufficing to 
produce the desired effect. This result is acquired when 
upon draining off the dye the section is found to have 
changed from the bright carmine to a deep violet tint. Wash 
the sections well in water, dehydrate by spirit, clear by clove 
oil, and mount in balsam. In this way the most beautiful 
effects are obtained, which are specially valuable and instruc- 
tive. The tint is much less fatiguing to the eye than the 
bright glare of carmine, whilst in the differentiation of 
nervous tissue in the hardened brain this method of stain- 
ing appears to me unrivalled. The deeper the tint desired 
the longer immersion in aniline must be practised, but for 
the more satisfactory and pleasing effects I have usuallj^ 
found it suffice to stain for half an hour in picro-carmine 
and fifteen minutes in aniline. 

Notes on the Reaction of the Dye. — This compound 



staining enliances tlie value of an ordinary picro-carminate 
preparation in the fact that a better differentiation of structure 
is obtained, and examination by artificial light rendered as 
agreeable and valuable as by day-light. With all the 
advantages of a picro-carmine staining we have conjoined 
tlie sjiecial action exhibited by aniline for the cell-processes 
and finer nervous meshwork, whilst at the same time the 
general tint is deeper than by picro-carmine, and the contrast 
betwixt the deeper and lighter stained parts more marked. 
The nem'oglia is stained faintly; the connective nuclei are 
prominent objects; the nerve-cells are well shown, the nucleus 
taking a much deeper tint. The axis-cylinder, in transverse 
sections of medullated fasciculi, is stained of a deep purple, 
the intertubular connective and its nuclei being of much 
lighter hue. The vascular channels and their nuclear elements, 
which are not satisfactorily shown in simple aniline staining, 
are exhibited to great advantage. I find this method of 
staining very suitable for displaying the coarse structure of 
the brain, and sections through a whole hemisphere treated 
by this method are most instructive for naked-eye examina- 
tion or by low-power objectives. 

2. Picro-Aniline Staining. — A useful staining of 
medulla and spinal cord may be obtained by emplojdng 
Judson's " Cambridge blue " as the first pigment, washing 
oif superfluous dye, and then by momentary immersion in a 
saturated solution of picric acid the section acquires a bril- 
liant green, which exhibits structural details remarkably well. 
Since both pigments are soluble in alcohol, the sections should 
be very rapidly dehydrated, cleared by clove oil, and mounted 
in the usual manner. 

3. Osmic Acid and Picro-carmine. — The section is 
immersed in a solution of osmic acid (1 per cent.) carefully 
covered over so as to confine this very volatile reagent as far 
as possible to the tissues to be acted upon. "When the latter 
has acquired a dark-brown tint the section should be removed, 
well washed, and lightly stained with picro-carmine. 


Such sections should be mounted in glycerine or glycerine 
jelly. Sections through the hemispheres of the brain in 
small animals, or the pons and medulla may be conveniently 
treated by the osmic acid contained in a watch-glass covered 
by a small glass plate. The above is a most valuable method 
of staining. 

4. Haematoxylin with Aniline. — This, like the last, 
gives a genuine double staining. Sections are first stained 
by logwood according to the directions already given, and 
immersed for a fcic seconds only in the aqueous solution of 
aniline blue-black (O'o per cent). A solution of chloral 
hydrate (2 per cent.) will remove any excess of aniline 
staining. Wash the sections well, rapidly dehydrate by 
alcohol, and mount sec. art. This process is chiefly adapted 
for sections of cerebellmn, and its special value will be 
referred to later on. The above are all the methods of 
staining I have found to be of greatest value, but I append 
here two methods of double staining for the cortex of the 
cerebellum, advocated by Professor Stirling,^ of the merits 
of which I have not had personal experience. 

5. Eosin and Logwood. — Stain a section for a few 
seconds in a very dilute watery solution of eosin (1 part to 
1,500 of water) until it has a faint red colour, Grreat care 
must be taken not to over-stain. Wash with water, stain the 
section with logwood, and mount in dammar. 

6. Picro-carmine and Iodine Green. — Stain a section 
in picro-carmine. Wash it in water acidulated with dilute 
acetic acid, and, after washing in pure water, stain again by 
means of iodine green, and mount in dammar. 


The Cortex Cerebri. — The cortex at the vertex in the 
frontal regions should by preference be stained by the 

' "Practif-al Histolog'y," p. 100. 



aqueous solution of aniline blue-black which has so remark- 
able an affinity for the nervous protoplasmic masses and their 
extensions, and which in these regions are so numerous. 
Next in value to aniline are the pioro-carminate and logwood 
dyes. With care very beautifid sections may be obtained 
from this part of the brain stained with picro-aniline, but a 
good aniline blue-black surpasses all others in intrinsic merits 
and for purely anatomical details. When it is desired to 
examine critically the contents of the nerve-cells with regard 
to morbid change the picro-carminate gives equally good 
results with the aniline. No reagent, on the other hand, 
exhibits so remarkably the proliferation of Deiter's corpuscles 
in the cortex as does aniline. 

In more posterior planes, and especially at the occipital 
pole of the hemisphere, where layers of the granule-cell 
formation predominate, the hematoxylin dye brings out the 
distinctive features of these realms better than any other. 
The nuclei of the crowded granide series are deeply stained 
by this reagent, although the large nerve-cells in the inter- 
mediate layers are but faintly seen. It is on this account 
advisable to attempt double staining by logwood and 
aniline when both granule cells and the larger nerve-cells 
are well shoAvn. The same remarks apply to the cortex in 
the olfactory region, where, in like manner, small granule 
elements predominate. 

The peculiar crowded cell-formation of the cornu ammonis 
stained by aniline blue-black affords beautiful preparations. 

The Cortex Cerebelli. — The granule layer of the 
cerebellar cortex is readily stained by any of the pigments 
more generally used, but not so the layers of Pm-kinje or their 
cell-processes, which branch out so abundantly in the pure 
grey layer. 

Picro-carmine and logwood alike yield fine, yet imperfect, 
preparations, since the mere outline of the Pm-kinje cell or 
its nucleus is often all that is visible in this layer. Occa- 
sionally, however, a few branches may be seen. To exhibit 


the great profusion of branches arising from these cells stain 
with aniline black, or preferably employ the double staining 
by logwood and aniline black. In these cases the granule 
layer will be of a bright purplish blue, whilst the cells of 
Purkinje will be distinctly mapped out, the nucleus of a 
bluish black tint, and their branches displaying a perfect 
forest of twigs. 

The Central Medulla. — The student should make sec- 
tions, both in a vertical and horizontal direction, through 
the brain of the smaller mammalia, e.g., the cat, dog, or 
rabbit, and stain these preparations with the double object of 
utilizing them for coarse examination by the naked eye or 
two-inch objective, to learn the relationships of the various 
central structures at different planes, as also for the more 
arduous duty of tracing out the course taken by the different 
systems of meduUated fasciculi in the interior of the brain. 
Such preliminary work he will find invaluable when he 
attempts to unravel the intricate mechanism of the human 

For the coarse examination of the larger sections through 
the whole brain or one hemisphere, the aniline picro-car- 
minate has always appeared to me the better method of 
staining, but for the more minute microscopic tracing of 
central fasciculi the double staining by osmium and picro- 
carmine is the only reliable method. 

It must be remembered that whenever we are dealing with 
sections carried transversely to the axis of the medullated 
fasciculi these latter will be well delineated by aniline blue- 
black, picro-carmine, or logwood ; but where the plane of 
section lies parallel with the long axis of these fasciculi, the 
more reliable staining is that of the medullated sheath by 
means of osmic acid. Hence it arises that in following out 
the lengthened course of fibres through the brain by sections 
along their planes, osmium is the most valuable adjunct, 
whilst the structure of the medulla, studied in transverse 
section, is best treated by other reagents. Although when 


dealing witli the more extensive areas, as in sections tlirough 
the hemispheres, it is advisable to obtain extremely thin 
j)reparations, yet the vast majority of valuable sections for 
the purpose of tracing the central medulla are most unsuit- 
able for minute examination of the cortex, the details of 
which will be very poorly delineated. The choicest sections 
through the cortex, such, in fact, as are essential to a favour- 
able investigation of its structure, must be of extreme 
tenuity — a degree indeed not attainable in large sections 
through the hemispheric mass. 

Let the series of preparations illustrative of the minute 
structure of the cortex in diiferent realms, therefore, be 
wholly distinct from those illustrative of the central medidla 
and basal ganglia, since the methods of preparation which 
are most suitable for these diverse parts diifer so essentially. 
On the other hand, Exner's process is available for both 

The Pons, Medulla, and Spinal Cord. — The same 
method of staining is applicable to all these parts. Logwood 
has been used extensively for these regions, carmine still 
more so ; whilst the anihne reagent, which I regard as of far 
the greater value, has been comparatively neglected, at least 
so I judge from the fact that whenever sections of cord or 
medulla are exhibited, the carmine preparations infinitely 
outnimiber the aniline. A tolerably thick section of medulla 
or cord appears wonderfully thin when stained by carmine 
and cleared up by oil of cloves ; it shows to very fair 
advantage even to the higher powers of the microscope, and 
any inequality of surface or scoring from the frayed edge 
of the section-knife is obscured or entirely concealed from 
view. This is not the case with the aniline preparations. 
The depth of tint here not only warns us but too surely of 
any undue thickness of our section, but shows up any faults 
in the section due to scoring or tearing of structiu-e to a 
remarkable and most unpleasant degree. Hence, I incline 
to the belief that faults really attributable to the section- 


cutter and liis blade' have been credited to the dye, which 
has, therefore, fallen into comparative disrepute. Of this, 
however, we may be certain, that sections of medulla and 
cord properly stained by aniline blue-black are the most 
beautifvd preparations of these regions we can by our present 
methods possess ; and that a really good aniline preparation 
of these parts must of necessity be a reyy thin and perfect 
section, — a statement which can scarcely be made of carmine 
preparations. The student should therefore possess himself 
of the thinnest possible sections of the different regions of 
the medulla and sjjinal cord, prepared by the aqueous 
solution of aniline blue-black, and with due care to the con- 
dition of his section-knife, he may thus prepare a most 
invaluable series of sections ; yet from the necessary fine- 
ness of the sections, he will meet with greater difficulty in 
obtaining perfect preparations than by the carmine staining. 
Whilst thus giving the foremost place to aniline blue-black 
for staining these regions, I would not wish to undervalue 
the merits of picro-carmine. Next to aniline, it is pre- 
eminently useful here. Do not, however, be deceived by its 
peculiar power of concealing the demerits of your section, 
and so be led to regard a section as thin because it appears 
so when mounted. In this way, minute details may be 
wholly overlooked which really thin sections would have 
exhibited. With the high powers the actual thickness of 
the section may be decided readily. 

Apparatus required for Staining and Mounting. 

Bottles of staining reagents 

Porcelain capsules 




Tray for fresh slides 

Bottles of mounting media 

Grlass rods 


Camel-hair brush 

Funnel and filtering paper 

Mounted needles 


Bottles of cement 



Among the facilities offered for immediate examination of 
brain and spinal coi-d in the recent state, two processes 
present themselves for examination ; these are — 

1. Methods of section-cutting and preparation of frozen 


2. Methods of dissociation and teasing out of fresh tissues. 

We should perfectly familiarize ourselves with all these 
methods. There are certain difficulties peculiar to the pre- 
paration of fresh nervous tissues, and not met with in the 
manipulation of hardened sections, with which it is well to 
be acquainted. We shall recognize in the details of pre- 
paration now to be given, the means adopted for overcoming 
these obstacles. 

Sections of frozen brain must be extremely thin for satis- 
factory preparations, and these most delicate films absorb 
water so rapidly that the constituents undergo rapid dissocia- 
tion when floating in water. Again, after removal to a slide, 
the upper surface is found highly repellant of watery 
solutions, so that the application of osmic acid equably over 
its surface is a matter of some difficulty when dealing with 
so fine a film. Lastly, fine sections of spinal cord exhibit 
such rapid endosmose of fluid, that unless each section be 
removed immediately it is floated off the blade into glyce- 
rine, the nervous elements swell up and burst through the 
encircling girdle of membranes, becoming completely dis- 


sociated. The first difSculty as regards the cortex is met by 
the osmio acid treatment, the second is overcome by a 
little i^ractical manipulation, and the difficulty connected 
with section of the cord is wholly avoided by glycerine 
diluted with one-half or two-thirds its bulk of water. 

It must be remembered that all alcoholic and highly astringent 
reagents must be excluded from our list for staining purposes, 
and therefore haematoxylin and several other dyes employed 
for hardened brain are wholly inadmissible for fresh nervous 
tissues. We are, therefore, chiefly restricted to aniline blue- 
black, carmine, picro-carmine, and osmic acid. The osmium 
and aniline are especially valuable agents. 


The Freezing Method. — Use the ether freezing micro- 
tome and secure it to a firm support, as a table or ledge in 
front of an open window, where a cool di-aught plays over 
the section-plate. Place a large deep glass vessel half -filled 
with clear water on the support in front of the microtome ; 
have the ether-spray apparatus ready on the left hand, and 
the section-blade on the right. Throw across the knees a 
soft towel, with which the under surface of the section-blade 
has to be gently wiped occasionally to remove superfluous 
moisture. Next take up the section-blade, and holding it 
horizontally, pour upon its upper surface a small quantity of 
ether, allowing it to sweep from end to end, and agitate the 
blade in water until the blade, on removal, remains perfectly 
wet from end to end. Repeat the ether wash if the surface 
still remains somewhat greasy, as in this state it is unsuited 
for section-cutting. Lower the freezing chamber of the 
microtome until its surface, or freezing plate, is four or six 
millimeters below the upper surface of the section-plate. 
Place upon the upper surface of the freezing chamber the 
tissue to be frozen, which should be but a little over a quarter 


of an inch thick. Introduce the nozzle of the ether-spray 
instrument through the opening in the left-hand of the 
freezing chamber, so that the spray be directed upwards 
against the freezing plate immediately beneath the tissue. 
A steady current will, in a few seconds, cause blanching of 
the tissue next to the plate, to which it now firmly adheres, 
and fi'eezing proceeds rapidly upwards, as evidenced by its 
colour and firmness. It is not necessary to freeze above the 
level of the section-plate. Remove the unfrozen tissue down 
to a level with the section-plate by a sweep of the blade — 
plunge the latter into water, lightly wipe its iinckr surface 
upon the towel, and raising the section by the micrometer- 
screw, cut the first section with a sweep away from you, and 
from your left to yom* right hand. The section must be 
floated off in the glass vessel, and not plunged beneath the 
sm^face of the water. 

One should aim at acquii'ing the greatest expedition at this 
work, rapidly floating off each section, and wiping the blade 
preparatory to the next. It is all-important that fresh 
sections be removed from the water in which they float as 
quickly as possible, whilst, at the same time, a number of 
sections should be cut before the frozen mass begins to thaw and 
loosen from the plate. Practised manipulators can with ease 
cut a dozen fine sections ere the mass loosens from the plate, 
but the tyro, Avhen simply practising the cutting of sections, 
is slow in his manipidation, and finds the mass loosens before 
he has secured three or four specimens. Two or three puffs 
from the spray, however, will, at this stage, fix it to the plate 
again, and thus he may with ease cut through the whole depth 
of tissue with very little expenditm-e of ether. When he has 
obtained sufficient manipulative skill at section-cutting, he 
will find it advisable to pass the first half-dozen sections 
through the second stage of the process before he cuts another 
series, so as to avoid the injury resulting from prolonged 
floating in water ; this last statement holds good for all 
nervous tissues alike. There is, perhaps, no method of 
obtaining fine sections of tissue so exj)editiously and with 


such extreme simplicity of detail as that just described, and 
certainlj none involving so little expenditure of time or labour ; 
yet, as the beginner will always meet with some obstacles in 
his first attempts, however simple the process, it will be as 
well to indicate those which most frequently present them- 
selves. His failiu'ss will invariably be due to one or other of 
the following causes : — 

1. The 1 0)12)6 rature of the room above 60° F. This also 
involves unnecessary waste of ether. Freezing should only 
be practised by ether spray when the temperatm'e of the 
atmosphere falls below 60° F. 

2. The brain mass frozen too high. Brain-tissue, when 
fi'ozen hard, cannot be properly cut ; in fact, our sections are 
invaiiably obtained from the tissue which is slowly thawing, 
and which is always of most delicate consistence for cutting. 
On this account it is not necessary to freeze up to the level of 
the section-plate. 

3. Too much uxiter on the blade. If this superfluous fluid 
runs off on to the section and freezing-plate, it sets into a 
solid icy mass, which injm-es the blade and greatly retards 

4. Water freezes over the. section-dilate. The uneven sur- 
face thus produced prevents section-cutting. The difficulty 
is removed by never neglecting to draw the under surface of 
the blade over the towel before each section is taken. 

5. Brain freezes too Jiard inrspcctive of the above conditions. 
This only results when the tissue is extremely oedematous 
from morbid states, and when the temperature of the siu'- 
rounding atmosphere is so low that freezing is unusually 
rapid. The difficulty is met by moistening the upper siu'face 
of the blade with glycerine instead of water, and carefully 
guarding it from extending to the freezing plate. 

6. Sections tear or cling to the blade. This residts from 
the greasy surface of the blade retaining no water to float up 
the section. 

7. Tissue cannot be frozen to plate or loosens too soon. The 


first occurs when glycerine or other medium which cannot be 
frozen has been accidentally smeared over the plate — the 
latter usually from want of rapid manipulation on the part of 
the operator, or too warm an atmosphere for satisfactory 

8. Lastly, the icell or freezing chamber may he too sni'dl, 
in which case freezing is not only retarded, but subsequent 
thawing is hastened. For my own part I would strongly 
recommend this portion of the instrument to be full 2 inches 
in diameter by 1| inches deep. 

We now proceed to the subsequent treatment of our 

Staining Fresh Sections of Cortex. — In this stage our 
sections pass through two processes — the osmic acid treat- 
ment and aniline staining. Take a perfectly clean glass 
slide, pass it beneath a section floating in water, and, fixing it 
to the slide by a needle, remove it carefully, draining off all 
superfluous fluid from the slide. Remove each section in a 
similar way, and next, by means of a pipette, let two or three 
drops of osmic acid solution (0"25 per cent.) fall on the slide 
and float up the section ; with a pen-knife carefully spread 
this fluid over the upper surface of the section, which, on 
account of the repulsion afforded, gives a little trouble with 
extremely thin films. This procedure is, of course, not 
necessary where the section has been sunk in water, but the 
more delicate sections float, and can be plunged beneath the 
surface of the water only at the risk of ruining them. A 
little practice soon enables us to carry the osmic acid over the 
most delicate films without tearing them. Sections thus 
treated should not remain over one minute in the osmic acid 
of the strength named, but should at once be immersed in a 
vessel of pure water and left here for five or ten minutes. By 
this time all superfliious acid has been removed, . and the 
osmium has " fixed " the nervous structures, so that they are 
manipulated without danger and do not deteriorate in water. 
The sections may now be stained collectively, or, as I prefer 


it, each section should be placed on a slide, and sufficient 
aniline blue-black dye (0'25 per cent.) poured on to completely 
cover the film. Place the slides under cover. It is well to 
have a small tray with a shelf attached around its sides on 
which the slides may rest, the tray being fitted with a cover. 
The staining with the aqueous solution of the dye occupies 
about one hour ; alcoholic solutions of the dye are of course 
inadmissible. When staining is complete a pipette will 
remove superfluous dye, or it may be drained off carefull}'', 
and the sections again plunged into water and gently moved 
about imtil well washed from unfixed dye. There are certain 
circumstances which the student shoidd be aware of in adopt- 
ing this process of staining, and these I will here briefly 
allude to. 

If the osmic acid solution be too strong or left on the 
section too long, subsequent staining by aniline will prove 
very unsatisfactory, and, moreover, the osmium staining 
becomes apparent, and is dissolved out subsequently by the 
benzole solution of balsam tinting the latter strongly and 
spoiling the preparation. Since the osmic acid is used simply 
to give the sections the necessary consistence for manipulation, 
and not for the purpose of staining, it should be employed in 
the weakest form consistent with this action, and the sections 
exposed to its agency for the very shortest period necessary, 
and very thoroughly washed prior to staining. If the osmic 
acid be unequally spread over the section, those parts not 
immersed will stain with less vigour by aniline. 

Again, when all the above precautions have been taken, it 
is sometimes found that sections exhibit a rough, irregularly- 
stained surface, really due to the edge of the section-blade 
being somewhat frayed, and so tearing the delicate structures 
in cutting through them. 

Mounting for Permanent Preparations. — Let each 
section, when well washed, be received upon the centre of a 
perfectly clean glass slide ; all fluid drained off, and the 
specimens again placed upon the shelf of the covered tray, to 


dry spontaneously. The process may be hastened, if neces- 
sary, by enclosing them within a bell-glass along with a 
capsiile containing strong sulphuric acid. When perfectly 
dehydrated, let a drop or two of a benzole solution of balsam 
fall on the section, and adjust the cover- glass. 

Staining Fresh Sections of Central Medulla. — 

As before stated, the same method of staining is not alike 
suitable for both cortex and medulla. The aniline process, 
just described, is in every way adapted for the cortex of the 
cerebrum and cerebellum ; but when our object is to follow 
the com-se of the central medullated fasciculi, and examine 
the structiu'e of the basal ganglia, the use of osmic acid has 
to be invoked. Before practising on the more extensive areas 
of the human cerebrum, it would be well for the student to 
slice the brain of one of the smaller animals — (the cat, dog, 
rabbit, are very suitable) — upon the freezing microtome, 
carrying sections through the hemispheres in vertical and 
horizontal planes. Such sections should be extremely fine, 
and on removal on a slide to the stage of the microscope, 
shoidd exhibit, in an exquisite manner, the details of 
medullated structures. The grey matter will, of course, be 
translucent, and apparently structureless ; but the medullated 
fasciculi will appear in striking relief of a pure white, the 
minutest fibre readily followed by a comparatively low-power 
objective. Our object, however, is to render these structures, 
not only clearly-defined, but permanent preparations, and this 
may be effected by the following procedure. The finest 
sections are removed as rapidly as possible from the water in 
which they float upon a slide, and floated up immediately by 
osmic acid (1 per cent, solution)^ drawn also carefully over 
the upper surface of the section. The slide is placed aside, 
closely covered over, so as to confine the vapom- of this very 
volatile reagent as much as possible to the section. In a 
period, varying from five to ten minutes, the sections are 
floated off into Avater, and freely washed. They acquire 
rapidly the deep brown tint of osmium, and can be manipu- 


lated without fear of tearing. The depth of tint required 
can be best learnt by experience. 

After well washing, the sections may either be mounted as 
they are, or immersed for a few moments in picro-carmine ; 
the latter is, perhaps, the best plan to adopt, as the grey 
matter is thus shown stained, in striking contrast to the 

The mounting of these preparations is more tedious, as 
they require to be j)reserved in glycerine or aqueous solutions, 
and careful sealing-up is therefore requisite. Take up the 
sections on the centre of a slide, after washing off the picro- 
carmine dye, and allow a drop or two of strong glycerine to 
float up the sections and remain exposed for five or ten 
minutes. The section becomes clearer and freed from much 
water by this means, and the glycerine is drained off. Just 
sufficient strong glycerine is again di'opped on to the section 
to cover it completely, and the cover is placed on. All traces 
of superfluous glycerine must be carefully removed from the 
margins of the cover-glass after it has been gently pressed 
down, and on no account must any glycerine remain on the 
slide just outside the cover- glass. A thin coating of the lead 
cement is laid on around the square or circle of thin glass, 
just overlapping by a couple of millimeters the edges of the 
latter. This must be allowed to dry completely, when a 
second coating is run on, and also allowed to dry. Over this, 
a layer of gold size may be painted, and a finish afforded by 
the Avhite zinc paint. This method of sealing-up, although 
tedious, is most reliable and durable ; it well repays the 
trouble taken. The same method may be employed where 
preservative fluids, such as Farrant's, Groadby's, or sodium 
chloride with camphor solution are - used. We should be 
cautious in using these fluid preservatives, to lay the gold 
size above and not beneath the red lead cement. A weak solu- 
tion of chloride of sodimn, made with camphor- water, also 
answers well for preserving the preparations stained by osmie 
acid alone. 




Prior to the systematic section-cutting of frozen brain, 
various methods of fresh preparation had been tried. Thus, 
a thin section of cortex was often pressed out between the 
cover-glass and slide, and its vascular system fairly well 
displayed ; but the nerve-cells and their delicate ramifications 
could not, of course, be detected by such a coarse method of 
procedure. It was found necessary to prepare the section by 
certain reagents whereby dissociation of elements might be 
secured, whilst some form of staining reagent was likewise 
essential. Amongst the earher attempts of this kind, was 
that made by Grerlach,^ but the process is applicable only to 
the large ganglionic cells of the spinal cord. I have myself 
employed a process less tedious than that of Grerlach, having 
the advantage of being rapid and immediate in application, 
with results which, to me, appear equivalent to what is 
obtained by the more lengthened process. I shall describe 

Gerlach's Method of Dissociation. — The finest pos- 
sible sections are made longitudinally, through the perfectly 
fresh spinal cord of the calf or ox, and are immediately placed 
in a very weak solution of bichromate of ammonia (1 part to 
5,000 or 10,000 parts of water). Let them remain here for 
two or three days, at a cool temperature. Next, immerse 
them for twenty-four hours in a dilute solution of ammoniacal 
carmine. Wash the sections well with distilled water, and 
tease out the nerve-cells with a needle, under a lens. Preserve 
in glycerine, or evaporate to dryness on a slide ; moisten with 
oil of cloves, and mount in Canada balsam. 

Dissociation may thus be practised in the case of ganglionic 
cells, wherever they are found, the macerating fluid being 
either dilute bichromate of potash or ammonia, Miiller's fluid, 

' Strieker's " Human and Comparative Histology." Syd. Soc. Vol. ii. 
p 345. 


or iodized serum, the process of maceration occupying from 
one to three days. 

Rapid Dissociation and Teasing. — Make a fine 
section with a razor through the anterior cornu of the 
cord in its lumbar enlargement. The section must be 
made longitudinally, and either by hand or on the freezing 
microtome. Place the section in the centre of a glass slide ; 
saturate it by immersion in Miiller's fluid for a few minutes ; 
place a glass cover over the section, and gradually compress 
it by a mounted needle between the cover-glass and slide. 
Before a fine film is obtained, the shde is carefully wiped on 
its under sm-face, and examined by a low power under the 
dissecting microscope. Even as opaque objects, the ganglion 
cells Avill now attract our attention by their little masses of 
pigment. With the slide still under a low objective, steadily 
and gradually compress the part where the cells chieflj' con- 
gregate, and, by a gentle rocking movement of the cover- 
glass, we may thus spread them out over the field, or aggre- 
gate towards the clearer areas, whilst any alteration they 
might suffer from this compression, is sure to be very trivial, 
owing to the elastic resistance of the neuroglia framework. 
The cover-glass may now be partly pressed and partly tilted 
off the film, and the mounted needle used to arrange the cells 
according to our mind, whilst the other textures around may 
be carefully removed. A few drops of Beale's strong 
solution of carmine, or the aqueous solution of aniline blue- 
black (0'25 per cent.), will very rapidly tint the germinal 
centres of the required depth of hue, and should then be 
removed by immersion in water, or by allowing a few drops 
of water gently to flow over the preparation. The cells are 
dried in situ, moistened with clove oil, and mounted in 

Dissociation and Staining of Ganglion-cells of 
Cortex. — The two methods just detailed are available 
only for the cells of the spinal cord. For the ganglion- 
cells of the human cortex, I devised a method which 


was described in the Monthly Microscopical Journal for 
1876, and which affords very satisfactory results. The 
process may be considered under three stages: 1. Obtaining 
the fihn ; 2. Staining ; 3. Permanent mounting of the prepa- 

1. First Stage: The Film. — Strip the convolution of 
its membranes, and excise a portion convenient for manipida- 
tion. Hold it betwixt the thumb and second finger of the 
left hand, so that the index finger may guide and support the 
blade. Then, with the razor-blade satiu-ated with methylated 
spirit, cut section after section vertically to the surface of the 
cortex, and of the greatest delicacy possible by these means. 
Very thin sections may thus be obtained with little trouble. 
Each should be placed on a glass slide, and three or four 
drops of Miiller's fluid allowed to fall upon them from a 
pipette. Maceration in this fluid for a few seconds, prevents 
the film adhering, dming the next procedure, to the cover- 
glass or slide. The cover is next placed on, and so arranged 
as to cover the film by only one-half its diameter. Place the 
point of a mounted needle upon the centre of the cover-glass, 
and by steady pressure flatten out the section into an 
almost transparent film. Practice will soon enable the 
operator to so arrange the cover-glass that the extreme edge 
of the compressed section still occupies the space covered by 
the glass circle. Now remove the superfluous reagent by 
rapid rinsing in water, and place the slide in a flat porcelain 
bath or tray, containing methylated spirit. In from thirty 
to forty seconds, the cover can be removed without tearing 
the film. For this purpose, remove the slide, steady the cover 
by the pressure of a finger against one part of its edge, 
and pass the point of a needle under the edge, near the 
medullated portion of the section, and gradually elevate 
it. "Wash the films from spirit by allowing a stream of water 
to flow over them from a large camel-hair brush. 

2. Second Stage : Staining. — The aniline blue-black 
solution (1 per cent.), is the most suitable staining reagent. 


It is dropped upon the film as it lies oil the slide, and when 
sufficiently stained, the superfluous dye is poured off, and the 
slide lowered gently into water, the film being washed by 
agitating the fluid around it with a camel-hair brush. Car- 
mine and picro-carmine may be also used for staining ; but 
hithei'to the best results have been obtained with aniline black 

3. Third Stage : Permanent Mounting. — The speci- 
mens lying in the centre of their respective slides are now 
transferred to a bell-glass, so as to exclude dust, and left to 
dry spontaneously, or the process of desiccation is hastened by 
including a capsule containing strong sulphm'ic acid. When 
perfectly dry, let a drop of chloroform fall on the preparation, 
and immmediately add the balsam (chloroform or benzole 
solution), and adjust the cover glass. 

Examination of Neuroglia. 

The neuroglia may be well shown in the spinal cord by 
the methods of dissociation above given. It has been especi- 
ally studied in the fresh state by Boll, Eanvier, Mierzejewski, 
and others, and the following methods are worthy of note : — 

1. Boll's Method. — Thin sections of the brain-substance 
were plunged into a solution of osmic acid, O'l per cent., and 
after twenty-four hovu's immersion, were washed in distilled 
water and kept in a concentrated solution of potassic acetate. 
For microscopic examination, .these darkened sections were 
dissociated or compressed strongly betwixt cover and slide. 

2. Ranvier's Method. — Osmic acid (1 part in 300 parts 
of water) was injected interstitiaUy into the fresh spinal cord. 
The pai-ts infiltrated by the reagent were afterwards removed 
by the section-knife, torn up, and teased in distilled water, 
coloured by picro-cai-mine, and examined in glycerine. Tliis, 
I may add, is a very excellent and valuable method. 

L 2 


3. Mierzejewski's Method. — ^A fragment of the white 
substance of the brain, about one cubic centimeter in bulk, 
was placed for twelve to twenty-four hours in a solution 
of osmic acid (1 part in 300 parts of water). By this time 
the preparation was impregnated by the reagent and its 
exterior dark and callous. The hardened exterior was then 
sliced off by the section-knife, when the parts immediately 
beneath it were found pretty equably affected by the osmium. 
Minute preparations were then obtained from this region, 
after the manner of Eanvier, viz., by staining with picro- 
carmine and examining in glycerine. Mierzejewski employed 
this method satisfactorily in his studies of the neuroglia of 
the brain in general paralysis.^ 

Dissection of MeduUated Strands in the Fresh 
Brain. — Much valuable information may be gleaned by the 
student in the examinatian of the brain of the smaller 
mammalia by dissection beneath a fixed lens or dissecting 
microscope. The brain should be placed beneath the light 
concentrated by a bull's-eye condenser. The scalpel, 
mounted needle, and camel-hair pencil are all the aids he 
requires. In fact, the needle and brush do nearly all the 
work, the broken-down tissue and grey matter being washed 
off by the brush filled -with a solution of chloride of 
sodium (1 per cent.). It is astonishing to one who has not 
tried this method how much can thus be done. It is well to 
begin with the very small brains, such as that of the rat. 

Remarks upon Dissociation of Nerve-Cells. — The 
student must not regard the methods of dissociation just 
given as being merely a rough and ready means of displaying 
the coarse structure of nervous tissue. Where the microtome 
is not at hand, or cannot be employed, these methods are very 
valuable ; but, even when possessed of the permanent sections 
obtained by the microtome, dissociation should supplement 
our inquiries. The films thus obtained, as well as those off 

1 ' ' Etudes sur les Lesions Cerebrales dans la Paralysie Generale.' ' Archives 
de Fhysiologie, 1873. 


the freezing microtome, afford us miicli information upon 
histo- chemical examination. The elements may thus be 
treated by osmic acid, salts of gold, platinum, and silver, and 
a variety of dyes. For examination of the reactions with 
chemical reagents, I have found it useful to obtain the 
faintest tint by aniline, just sufficient, indeed, to bring into 
view the outline of the cells and their nuclei. For this pur- 
pose a few drops of very weak aniline dye, left on for a few 
minutes, and then washed off, may be fixed in the germinal 
centres by a di-op of extremely dilute hydrochloric acid, 
which is immediately cleared off by water, and the film 
left in a satisfactory state for noting the effect of reagents. 



The following list comprises fill the more useful reagents 
which the student will require in the prosecution of the 
minute examination of the brain. Those marked with an 
asterisk (*) are the most important, and cannot be dispensed 

Hardening Reagents. 

1.* Chromic acid (stock solution, 1 per cent.). 

Chromic acid cry st. ... 10 grammes. 
Water 1,000 c.c. 

Dissolve. Dilute to O'lo pei- cent, for brain, and 0-25 
per cent, for spinal cord. 

2.* Bichromate of potassium (stock solution, 4 per cent.) . 

Bichromate of potassiimi ... 40 grammes. 
"Water (hot) .;. ... 1,000 c.c. 

Dissolve. Used in dilutions from 0-25 per cent, upwards. 

3. Bichromate of ammoniimi (stock solution, 4 per 
cent.), made as the potassium solution above. 

4.* Mailer's fluid. 

Bichromate of potassium ... 25 grammes. 

Sodium sulphate ... ... 10 „ 

Water ... 1,000 c.c. 



Picric acid solution. 
Cold satiu-ated solution in water. 


6.* Osmic acid. 
A solution of 1 per cent, should be kept in a blue glass 
closely-stoppered bottle, to be diluted when required imme- 
diately before use. 

7.* Methylated spirit. 

8.* Absolute alcohol. 

Formula of tSi-Eci \.l Holutkjns. 

9. llutherford's reagent. 

Cliromic acid ... ... 1 gramme. 

Bichromate of potassium ... 2 ,, 

Water 1,200 c.c. 


10. Solutions for Betz's method. 

{a) Iodized alcohol. 

Alcohol (70 to 80 per cent.), tinted sherry- 
brown by tinct. iodi. co. 
{b) Iodized spirit. 

Methylated spirit, tinted sherry - brown by 
tinct. iodi. co. 
ic) Bichromate solutions. 

Solutions of potassium bichromate from 0*5 
per cent, up to 5 per cent. 

11. Solutions for Hamilton's method. 

(r/) Midler's fluid 3 parts. 

Methylated spiiit ... ... 1 „ 

[h] Solutions (aqueous) of ammonium bichromate, 

0"25 per cent, to 2 per cent. 

12. Kleinenberg's picric acid solution. 

Cold saturated aqueous solution of 

picric acid ... ... ... 800 c.c. 

Concentrated sulphmic acid ... 6 c.c. 

Mix, filter, and add to the filtrate, Avater 900 c.c. 


Staining Reagents. 

The following may be regarded as a fairly complete list of 
the reagents more generally employed ; the f ormnlie for special 
solutions are also given below : — 

1.* Carmine. ' Aniline dyes comprising — 

2* Hsematoxylin (crystals). irv* a -t it it ^ 

-r 1 . / 10.* Aniline blue-black. 

LiOgwood extract. -. , , , 

8.* Picric acid (crystals). ^' ,^ " .,. " , ^ 

. ^ "^ I -'-■^- Iv-osanihn (magenta). 

13. Fuchsine. 

14. Metli34-aniline. 

15. Iodine green. 

16. Judson's dyes, espe- 

cially *" Cambridge 

17. Various combinations 

5. Gold chloride. 
(J. Double chloride of gold 
and potassium. 

7. Silver nitrate (0'5 per 


8. Silver lactate (Alferow's 

formula) . , " p i i v 

. * ^ • • ■. 1 i \ of the above. 

d. Osmic acid (1 per cent.). ' 

Formula for Special Solutions. 

1.* Beale's carmine reagent. 

Carmine (in small fragments) 10 grains. 
Liquor ammonife ... • • ■ h drachm. 

Glycerine (Price's) 2 ounces. 

Alcohol ... •■• ••• ^ 5J 

Distilled water ... ••• 2 ,, 

Heat the carmine with the ammonia in a test-tube carefully, 
boil for a few minutes, and expose for one hour. Add the 
water ; filter. Add the spirit and glycerine, and expose until 
the odour of ammonia is scarcely perceptible. 
2. Thiersch's carmine reagent. 

(a) Cannine ••• ••• ••• 1 \)ia:i. 

Liquor ammonite ... • • • 1 ?> 

Distilled water 1 » 

Dissolve and filter. 

{h) Oxalic acid 1 P'^i't- 

Distilled water 22 „ 


Mix one part of (a) witli eight parts of (b), and add 
twelve parts of alcohol. 

3. Borax carmine. 

Carmine ... ... ... 1 part. 

Borax .., ... ... ... 4 „ 

AVater 56 „ 

Alcohol ... ... ... q.s. 

Dissolve the borax in the water, and add the carmine ; 
filter, and add to the filtrate two volumes of absolute alcohol. 

4.* Ranvier's picro-carmine. 

(a) Carmine 

Liquor ammonite . . . 

1 gramme. 
3 c.c. 
10 c.c. 

liub the carmine up with water ; add the ammonia, and 
dissolve with heat carefully applied, 

(b) Saturated aqueous solution of picric acid, 200 c.c. 

Add, when perfectly cold, (a) to (b), and evaporate with 
gentle warmth to one-third its bulk. Filter, and keep in 
stoppered bottle. 

5.* Minot's hsematoxylin dye. 

(«) Heematoxylin crystals 
Absolute alcohol 

Dissolve the hsematoxylin in alcohol, and add the alum, 
dissolved in the water. 

(b) Alum 1 part. 

Water ... 200 










Dissolve. For use, pour a little of (b) into a watch-glass, 
and add sufficient of (a) to colour it a light violet tint. 
Filter prior to staining. 


6.* Kleinenberg's hoematoxylin dye. 

(a) Make a saturated solution of crystallized calcium 
cliloride in alcohol (70 per cent.), and add alum 
to saturation. 

(b) Make a saturated solution of alum in alcohol (70 
per cent.), and add solution {a) to {h), in the pro- 
portion of 1 to 8. 

(c) To this resultant mixture, now add a feiv drops of a 
harely alkaline saturated solution of hsematoxylin. 

7.* Aniline blue-hlack (aqueous solution). 

Aniline blue-black ... 1 gramme. 
Distilled water 400 c.c. 

Dissolve, filter, and label " 0-25 per cent." Or, make a 
1 per cent, solution ; dilute and filter when required. 

8. Aniline blue-black (alcoholic solution). 

Aniline blue-black ... 1 decigramme. 

Water 4 c.c. 

Dissolve and add recti- 
fied spiiit ... 100 c.c. 
Filter, and label " O'l per cent." 

9. Aniline blue reagent. 

Aniline blue J grain. 

Distilled water 1 ounce. 

Alcohol 25 drops. 

10. Rutherford's rosanilin dye. 

Rosanilin or magenta (crystals) 1 grain. 

Absolute alcohol 100 minims. 

Distilled water 5 ounces. 

11. Dyes used in combination. 

Aniline blue-black (Aq. sol., 0-25 per cent.) 
+ ricro-carmine. 


Judson's " Cambridge blue " | 
+ Picric acid. j 

Osmic acid (1 per cent.) + Picro-carmine. 
Hsematoxylin ... + Aniline blue-black 

(0*5 per cent.). 
Eosin (1 to 1,500) ... + Hsematoxylin. 
Picro-carmine ... + Iodine green. 

12. Alferow's lactate of silver : — 

Lactate of silver ... ... 1 part. 

Distilled water 800 „ 

Concentrated solution of lactic 

acid ... ... ... A few drops. 

Note. — " The free acid renders precipitation less easy, and 
the chloride and albuminate of silver are alone formed." 

13. Gerlach's gold and potassium solution.^ 

Double chloride of gold and 

potassium ... ... 1 part. 

Water, feeblv acidulated 

with hydrochloric acid.. 10,000 ,, 

Sections acquire a pale lilac colom' in this solution in from 
ten to twelve hours. 

Solutions for Macerating and Dissociating 


1. Iodized serum. 

Fresh amniotic fluid ... 100 c.c. 

Tincture of iodine ... ... 1 c.c. 

Carbolic acid ... ... 2 drops. 

For small quantities, use the aqueous humour from the eye 
of sheep or ox. Other useful fluids are — 

2. Glycerine and water. 

' For staining sections of spinal cord by this method, see Art. " The Spinal 
Cord," by Gerlach, in Strieker's " Human and Comparative Histoloo-y." 


3, Chloride of sodium solution (0*75 per cent.). 

4, Ammonium and potassium bichromate (1 part to 

10,000 of water). 

5, Midler's fluid, freely diluted. 

Mounting Media. 

1.* Griycerine (Price's) ; 2.* Glycerine jelly (Rimming- 
ton's) ; 3.* Canada balsam ; 4.* Dammar varnish ; 5. Far- 
rant's solution. 

3.* Benzole solution of balsam. 

Canada balsam evaporated in a water-bath to a brittle, 
resinous consistency, is dissolved in benzole until it is 
sufficiently fluid to drop slowly from a glass rod. 

4.* Dammar fluid. 

Gum dammar, gum mastic, 

of each ... ... ... h ounce. 

Benzole ... ... ... 3 ,, 

Dissolve the gum in the benzole and filter. 

5.* Farrant's solution. 

Glycerine, saturated solution of arsenious acid, gum- 
arabic, equal parts of each. 

Let the mixture stand for several weeks, frequently stirring; 
and when no more of the gum-arabic is dissolved, filter the 

1.* Mr. Kitton's cement for glycerine mounting. 

White lead, red lead, litharge in powder, equal paris. 

Mix well. Keep a stock of the mixture ready ground in a 

For use — rub up a little of the powder with turpen- 
tine, and add sufficient gold size to allow it to work readily 
with a brush. The first coat should always be perfectly dry 
before the application of a second. 


2.* Zinc-white cement. 

Griim dammar. . . ... ... 8 oimces. 

Zinc oxide ... . . ... 1 ,, 

Benzole ... ... ... 8 ,, 

Dissolve the gum in the benzole, add the zinc, and strain 
through muslin. 

3. Marine glue. 
4.* Gfold size. 
5. Brunswick black. 
6.* Balsam or dammar varnish. 
In addition to the list of reagents above given, the student 
sliould supply himself with a bottle of clove oil, glacial acetic 
acid, and solution of chloral hydrate, 2 per cent. 

I ^^ D E X. 

Absolute weight of brain, 61 
Acid, chromic, 105, 153 

osmic, 105 

picric, 154 
Alferow's sUver lactate, 158 
Amnioniiun bichromate, 104, 154 
AnUice blue -black, 157 

picro-carminate, 129 

staining, 125 
Apparatus, displacement, 65 

specific gravity, 63 

for hardening tissues and section- 
cutting, 114 

for staining and mounting, 135 
Arachnoid, examination of, 9 
Arteries, capacity of, 20 

dissection of, 15 

nutrient, 24 

tunics of, 18 


Base of brain, 8 

Beale's carmine fluid, 155 

Betz's method of hardening, 103 

hardening solutions of, 154 
Bichromate solutions, 154 
Blood-vessels, distribution of, 10 

capacity of, 20 

coats of, 18 

nutrient, 24 
Boll's method for neuroglia, 147 
Borax carmine, 156 
Broca's estimates of capacity, 59 
Bucknill's specific gravity test, 63 

Canada balsam 159 
Capacity, arterial, 20 

cranial, 57 
Carmine staining fluids, 155 

staining process, 120 
Cements, list of, 159 
Central medulla, 133 
Chrome hardening, 102 
Chromic acid, 105, 153 
Cicatricial tissue, 44 
Circle of Willis, 16 
Colour of cortex, 45 
Compound pigments, 128, 157 
Consistence, estimation of, 29 

reductions in, 31 

augmented, 43 
Convolutions of brain, 75 

synonyms of, 78 
Cortex cerebelli, staining, 132 

cerebri, staining, 131 

congestion of, 52 

ganglion-cells of, 145 
Cranial capacity, 57 


Dammar mounting medium, 159 
Danilewski's method for weight, 07 
Dissection of brain, 69 
Dissociation of nervous tissue, 144 
cells in cortex, 145 
rapid method of, 145 
Gerlach's method of, 144 
Dissociating reagents, 158 
Dura mater, examination of, 3 




Ecker's s3monyms, 78 
Embolic pluggiuyr of brain, 36 
Eosin staining iluid, 131 
Exner's osniic acid process, 105 

EaiTant's solution, 159 

Flowers' method for cranial capacity, 

Freezing methods, 137 

microtomes, ether, 92 — 95 
Eutherford's, 90, 95 
Williams', 98 
Fresh brtiin, preparation of, 136 
sections, staining, 140 


Ganglia of brain, staining, 142 
Ganylion-cells of cortex, 122, 145 
Gerlach's gold and potassium solution, 
method of dissociation, 144 
Gravimetric methods, 60 


Hajmatoxylin dye, 156 

staining, process of, 116 
Hamilton's methods of hardening, 104 

hardening solutions, 154 
Hardened brain, preparation of, 100 
Hardening by chrome salts, 102 

by osmium, 105 

reagents, list of, 153 

Imbedding mixtures, 108 

process of, 107 
Iodine green reagent, 131 
Iodized serum, 158 


Kitton's lead cement, 159 
Kleinenberg's hgematoxylin dye, 117 
picric acid solution, 154 

List of apparatus for section-cutting, 
for staining and mounting-, 135 
solutions for hardening, 153 
dissociating, 158 
mounting, 159 
staining, 155 
Logwood staining reagent, 155 


Macerating reagents, 158 
Magenta dye, 157 
Medulla oblongata, 134 
Medulla, staining of central, 133 
Membranes, examination of, 1 — 14 
Methods of preparation, 86 
Microtome, ether freezing, 92, 95 

imbedding, 89 

Ranvier's, 90 

Eoy's, 91 

Eutherford's, 90, 95 

Schiefferdecker's, 91 

Stirling's, 90 

Williams', 98 
Mierzejewski, method for neuroglia, 

Minot's hfematoxylin dye, 116 
Mounting fresh tissues, 143 

hardened tissues, 115 

media, 159 
Miiller's fluid, 153 

Neuroglia, examination of, 147 
hypertrophy of, 44 

Osmic acid, process of hardening by, 
staining by, 130 

Pallor of cortex, 46 

Paraffin mass for imbedding, 108 

cast of cranium, 58 
Physical properties of cerebral sub- 
stance, 26 
Pia mater, examination of, 9 

vessels of, 22 



Picric acid solutions, 153,154 
Picro- aniline staining, 130 
Picro-cannine reagent, 156 

staining, 124 
Pigments, compound, 12S 
Pons, staining of, 134 
Potassitun bichromate solutions, 1-3 3, 

Preparation of fresh tissues, 136 

of hardened tissues, 100 
Pressiu'e, residts of intra-cranial, 54 

intra-thoracic, 54 
Puncta vasculosa, 53 
Putrefactive changes, 31 


Quekett Club microtome, 98 


Ranvier's method for neuroglia, 147 

microtome, 90 

picro-carmine dye, 156 
Reaction of aniline blue-black, 126 

carmine, 122 

hfematoxylin , 119 

osmic acid, 128 

picro-carmine, 124 
Reagents, list of, 153 
Reference to photographs, 77 
Roy's microtome, 91 
Rutherford's hardening fluid, 154 

microtome, 90, 95 

rosanilin dye, 157 


Sankey's aniline solution, 125 

specific gravity test, 63 
Schieff erdecker' s mici'otome, 91 
Sclerosis of nervous centres, 44 
Section-blade, 112 
Section-cutting, 110, 137 

Sections, prei)aration of hardened, 115 

preparation of fresh, 140 
Silver lactate, 158 
Sinuses of skidl, 5 
Softening, indications of, 32 

ischsemic, 33 

yeUow, 37 

inflammatory, 38 

oedematous, 41 
Special regions, staining of, 131 
Specific gravity of brain, 62 
Staining reagents, list of, 155 

methods, 115, 140 
Stevenson's displacement apparatus, 

Stii-ling's microtome, 90 
Siunniary of hardening methods, 106 
Synonyms of convolutions, 78 

Thiersch's carmine dye, 155 
Topography of brain, superficial, 75 
Tuke, method of estimating cranial 
capacity by Dr. Hack, 57 

Vault of cerebrum, examination of, 4 
Venous sinuses, 5 
Volume of brain, 55 
Volumetric methods, 55 


Weight of brain, absolute, 61 

dift'erential, 67 

specific, 62 
Williams' microtome, 98 

Zinc cement, 160 

Zones, congested cortical, 52 

Pardon and Sons, Printers, Paternoster Row, London. 

Catalogue B] 

Loftdon, II, Ne%v Burlington Street 
October, 1887 








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Abercrombie's Medical Jurisprudence, 4 
Adams (W.) on Clubfoot, 11 

on Contraction of the Fingers, 11 

on Curvature of the Spine, 11 

Alexander's Displacements of the Uterus, 6 
Allan on Fever Nursing, 7 

Outlines of Infectious Diseases, 7 

Allingham on Diseases of the Rectum, 14 
Anatomical Remembrancer, 3 

Balfour's Diseases of the Heart and Aorta, 9 
Balkwill's Mechanical Dentistry, 12 
Barnes (R.) on Obstetric Operations, 5 

on Diseases of Women, 5 

Basil's Commoner Diseases and Accidents, 10 
Beale's Microscope in Medicine, 8 

Slight Ailments, 8 

Urinary and Renal Derangements, 14 

Bellamy's Surgical Anatomy, 3 

Bennet (J. H.) on the Mediterranean, 10 

on Pulmonary Consumption, 10 

Bentley and Trimen's Medicinal Plants, 7 
Bentley's Manual of Botany, 7 

Structural Botany, 7 

Systematic Botany, 7 

Bowlby's Surgical Pathology and Morbid Anatomy, 10 

Braune's Topographical Anatomy, 3 

Brodhurst's Anchylosis, 11 

■ — — Curvatures, &c., of the Spine, 11 

Orthopeedic Surgery, ii 

Bryant's Acute Intestinal Strangulation, 9 

Practice of Surgery, 11 

Bucknill and Tuke's Psychological Medicine, 3 
Buist's Vaccinia and Variola, 8 
Bulkley's Acne, 13 

Diseases of the Skin, 13 

Burdett's Cottage Hospitals, 4 

Pay Hospitals, 4 

Burton's Midwifery for Midwives, 6 
Butlin's Malignant Disease of the Larynx, 13 

Operative Surgery of Malignant Disease, 13 

Sarcoma and Carcinoma, 13 

Buzzard's Diseases of the Nervous System 9 

Peripheral Neuritis, 9 

Carpenter's Human Physiology, 4 
Cayley's Typhoid Fever, 8 

Charteris on Health Resorts, 10 

Practice of Medicine, 8 

Chavers' Diseases of India, 8 
Churchill's Face and Foot Deformities, 11 
Clouston's Lectures on Mental Diseases, 5 
Cobbold on Parasites, 13 

Coles' Dental Mechanics, i^ 
Cooper's Syphilis and Pseudo-Syphilis, 14 
Coulson on Diseases of the Bladder, 14 
Courty's Diseases of the Uterus, Ovaries, &c., 6 
Cripps' Diseases of the Rectum and Anus, 14 
CuUingworth's Manual of Nursing, 6 

Short Manual for Monthly Nurses, 6 

Curling's Diseases of the Testis, 14 
Dalby's Diseases and Injuries of the Ear, 12 
Day on Diseases of Children, 6 

on Headaches, 10 

Dobell's Lectures on Winter Cough, 8 

Loss of Weight, &c., 8 

Mont Dor^ Cure, 8 

Domville's Manual for Nurses, 6 
Doran's Gynaecological Operations, 6 
Down's Mental Affections of Childhood, 5 
Draper's Text Book of Medical Physics, 4 
Druitt's Surgeon's Vade-Mecum, 11 
Duncan on Diseases of Women, 5 

on Sterility in Woman, 5 

Dunglison's Medical Dictionary, 14 
East's Private Treatment of the Insane, 5 
Ebstein on Regimen in Gout, 9 

Ellis's Diseases of Children, 6 
Fagge's Principles and Practice of Medicine, 
Fayrer's Climate and P'evers of India, 7 
Fenwick's Chronic Atrophy of the Stomach, 8 

Medical Diagnosis, 8 

Outlines of Medical Treatment, 8 

Flint's Principles and Practice of Medicine, 7 

Flower's Diagrams of the Nerves, 3 

Fox's (C. B.) Examinations of Water, Air, and Food, 5 

Fox's (T.) Atlas of Skin Diseases, 13 

Freyer's Litholopaxy, 14 

Frey's Histology and Histo-Chemistry, 4 

Galabin's Diseases of Women, 6 

Manual of Midwifery, 5 

Gamgee's Treatment of Wounds and Fractures, 11 

Godlee's Atlas of Human Anatomy, 3 
Goodhart's Diseases of Children, 6 
Gorgas' Dental Medicine, 13 
Gowers' -Diseases of the Brain, g 

Diseases of the Spinal Cord, 9 

Manual of Diseases of Nervous System, 9 

Medical Ophthalmoscopy, 9 

Pseudo-Hypertrophic Muscular Paralysis, 9 

Granville on Gout, 9 

on Nerve Vibration and Excitation, 9 

Guy s Hospital Formulae, 2 

Reports, 2 

Habershon's Diseases of the Abdomen, 9 

Liver, 9 

, Stomach, 9 

Pneumogastric Nerve, 9 

Hambleton's What is Consumotion? 8 
Hardwicke's Medical Education, 14 
Harley on Diseases of the Liver, 9 

Inflammations of the Liver, 9 

Harris's Dentistry, 13 

Harrison's Surgical Disorders of the Urinary Organs, 13 

Hartridge's Refraction of the Eye, 12 

Harvey's Manuscript Lectures, 3 

Heath's Injuries and Diseases of the Jaws, 10 

Minor Surgery and Bandaging, 10 

Operative Surgery, 10 

Practical Anatomy, 3 

Surgical Diagnosis, 10 

Helm on Short and Long Sight, &c., 11 
Higgens' Ophthalmic Out-patient Practice, 11 
Hillis' Leprosy in British Guiana, 13 
Holden's Dissections, 3 
Human Osteology, 3 

Landmarks, 3 

Holmes' (G.) Vocal Physiology and Hygiene, 12 
Hood's (D. C. ) Diseases and their Commencement, 6 
Hood (P.) on Gout, Rheumatism, &c., 9 
Hooper's Physician's Vade-l\Iecum, 8 
Hutchinson's Clinical Surgery, n 

Pedigree of Disease, 11 

'■— Rare Diseases of the Skin, 13 

Hyde's Diseases of the Skin, 13 

lames (P.) on Sore Throat, 12 

Jessett's Cancer of the Mouth, &c., 13 

Johnson's Medical Lectures and Essays, 8 

Jones (C. H.) and Sieveking's Pathological Anatomy, 4 

Jones' (H. McN.) Diseases of the Ear and Pharynx, 12 

— — Atlas of Diseases of Membrana Tympani, 12 

Spinal Curvatures. 11 

Journal of British Dental Association, 2 

■ ■ Mental Science, 2 

King's Manual of Obstetrics, 5 

Lancereaux's Atlas of Pathological Anatomy, 4 

Lewis (Bevan) on the Human Brain, 4 

Liebreich's Atlas of Ophthalmoscopy, 12 

Liveing's Megrim, Sick Headache, &c., 9 

London Hospital Reports, 2 

Lfickes' Hospital Sisters and their Duties, 7 

Macdonald's (J. D.) Examination of Water and Air, 4 

Mackenzie on Diphtheria, 12 

on Diseases of the Throat and Nose, 12 

McLeod's Operative Surgery, 10 
MacMunn's Spectroscope in Medicine, 8 
IMacnamara's Diseases of the Eye, 11 

Bones and Joints, 11 

Marcet's Southern and Swiss Health-Resorts, 10 

Martin's Ambulance Lectures, 10 

Mayne's Medical Vocabulary, 14 

Middlesex Hospital Reports, 2 

Mitchell's Diseases of the Nervous System, 9 

Moore's Family Medicine for India, 7 

Health-Resorts for Tropical Invalids, 7 

— ^ Manual of the Diseases of India, 7 

Morris' (H.) Anatomy of the Joints, 3 

Morton's Spina Bifida, 10 

Mouat and Snell on Hospitals, 4 

Nettleship's Diseases of the Eye, 12 

Nunn's Cancer of the Breast, 13 

Ogston's Medical Jurisprudence, 4 

Ophthalmic (Royal London) Hospital Reports, 2 

Ophthalmological Society's Transactions, 2 

Oppert's Hospitals, Infirmaries, Dispensaries, &c. , 4 

Osborn on Diseases of the Testis, 13 

on Hydrocele, 13 

Owen's Materia Medica, 7 
Page's Injuries of the Spine, 11 
Parkes' Practical Hygiene, 5 
Pavy on Diabetes, 10 

Pavy on Food and Dietetics, 10 

[Continued on the next pagt. 

Index — contimied. 

Pharmaceutical Journal, 2 
Pharmacopoeia of the London Hospital, 7 
PhiUips' Materia Medica and Therapeutics, 7 
Pollock's Histology of the Eye and Eyelids, 12 
Porritt's Intra-Thoracic Effusion, 8 
Purcell on Cancer, 13 
Quinby's Notes on Dental Practice, 12 
Raye's Ambulance Handbook, 10 
Reynolds' (J. J.) Diseases of Women, 5 

Notes on Midwifery, 5 

Richardson's Mechanical Dentistry, 12 
Roberts' (C.) Manual of Anthropometry, 5 

Detection of Colour-BIindness, 5 

Roberts' (D. Lloyd) Practice of Midwifery, 5 
Robinson (Tom) on Eczema, 14 

— — on Syphilis, 14 

Robinson (W.l on Endemic Goitre or Thyreocele, 12 
Ross's Aphasia, 9 

Diseases of the Nervous System, 9 

Handbook of ditto, 9 

Routh's Infant Feeding, 7 

Royal College of Surgeons Museum Catalogues, 2 

Royle and Harley's Materia IMedica, 7 

St. Bartholomew's Hospital Catalogue, 2 

St. George's Hospital Reports, 2 

St. Thomas's Hospital Reports, 2 

Sansom's Valvular Disease of the Heart, 8 

Savage on the Female Pelvic Organs, 5 

.Schweigger on Squint, 12 

Sewill's Dental Anatomy, 12 

Sharkey's Spasm in Chronic Nerve Disease, 9 

Shore's Elementary Practical Biologj', 4 

Sieveking's Life Assurance, 14 

Simon's Public Health Reports, 4 

Smith's (E.) Clinical Studies, 6 

■ Diseases in Children, 6 

Wasting Diseases of Infants and Children, 6 

Smith's (J. Greig) Abdominal Surgery, 6 

Smith's (Henry) Surgery of the Rectum, 14 

Smith's (Heywood) Dysmenorrhoea, 5 

Smith (Priestley) on Glaucoma, 12 

Snell's Electro-Magnet in Ophthalmic Surgery, 12 

Snow's Clinical Notes on Cancer, 13 

Southam's Regional Surgery, 11 

Squire's Companion to the Pharmacopoeia, 7 

Pharmacopceias of London Hospitals, 7 

Steavenson's Electricity. 11 

Stills and Maisch's National Dispensatory, 7 

Stocken's Dental Materia Medica and Therapeutics, 13 

Sutton's General Pathology, 4 

Swain's Surgical Emergencies, 10 

Swayne's Obstetric Aphorisms, 6 

Taylor's Medical Jurisprudence, 4 

Taylor's Poisons in relation to Medical Jurisprudence, 4 

Teale's Dangers to Health, 5 

Thin's Cancerous Affections of the Skin, 13 

— Pathology and Treatment of Ringworm, 

Thomas's Diseases of Women, 6 
Thompson's (Sir H.) Calculous Disease, 14 

Diseases of the Prostate, 14 

— Diseases of theUrinarj' Organs, 14 

• Lithotomy and Lithotrity, 14 

Stricture of the Urethra, 14 

Suprapubic Operation, 14 

Surgery of the Urinary Organs, 14 

Tumours of the Bladder, 14 

Thorowgood on Asthma, 8 

on Materia Medica and Therapeutics, 7 

Thudichum's Pathology of the Urine, 14 
Tibbits' Medical and Surgical Electricity,- 10 

Map of Motor Points, 10 

How to use a Galvanic Battery, 10 

Electrical and Anatomical Demonstrations, 10 

Tilt's Change of Life, b 

Uterine Therapeutics, 6 

Tirard's Prescriber's Pharmacopoeia, 7 
Tomes' (C. S.) Dental Anatomy, 12 
Tomes' (J. and C. S.) Dental Surgery, 12 
Tuke's Influence of the Mind upon the Body, 5 

Sleep-Walking and Hypnotism, 5 

Vintras on the Mineral Waters, &c., of France, 10 
Virchow's Post-mortem Examinations, 4 
Walsham's Surgery : its Theory and Practice, 11 
Waring's Indian Bazaar Medicines, 7 

Practical Therapeutics, 7 

Warlomont's Animal Vaccination, 13 
Warner's Guide to Medical Case-Taking, 8 
Waters' (A. T. H.) Diseases of the Chest, 8 
Weaver's Pulmonary Consumption, 8 
Wells' (Spencer) Abdominal Tumours, 5 

Ovarian and Uterine Tumours, 5 

West and Duncan's Diseases of Women, 5 
West's (S.) How to Examine the Chest, 8 
Whittaker's Primer on the Urine, 14 
Wilks' Diseases of the Nervous System, 8 
Williams' (Roger) Influence ot Sex, 4 
Wilson's (.Sir E.) Anatomists' Vade-!Mecum, 3 
Wilson's (G.) Handbook of Hygiene, 5 

Healthy Life and Dwellings, 5 

Wilson's (W. S.) Ocean as a Health-Resort, 10 
Wolfe's Diseases and Injuries of the Eye, 11 
Year Book of Pharmacy, 2 
Yeo's (G. F.) Manual of Physiology, 4 
Yeo's (J. B.) Contagiousness of Pulmonary Consump.^ 
tlon, 8 

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